ES2833506T3 - Liquid pharmaceutical composition - Google Patents

Abstract

A liquid pharmaceutical composition comprising: from 20 to about 70 mg / ml of etanercept (including any biosimilars thereof); a 10 to 40 mM buffer system, the buffer system being a phosphate buffer system and / or a citrate buffer system; 20 to 150 mM sodium chloride; 0.5 to 5% by weight of sucrose; and either aspartic acid, or a combination of lysine and proline; wherein: the liquid pharmaceutical composition has a pH between 6.1 and 6.5; the liquid pharmaceutical composition is free of arginine or comprises arginine in a concentration of at most 0.1 mM; the liquid pharmaceutical composition is free of sulfur-containing amino acid (s) or comprises sulfur-containing amino acid (s) in a concentration of at most 0.1 mM; the liquid pharmaceutical product is free of nitrogen-containing chelating agent (s) or comprises nitrogen-containing chelating agent (s) in a concentration of not more than 0.1 mM; and the liquid pharmaceutical composition containing a combination of lysine and proline contains 3.5 to 4.5% by weight of sucrose.

Description

DESCRIPTION

Liquid pharmaceutical composition

Introduction

The present invention relates to a new liquid protein formulation. In particular, the invention relates to a liquid pharmaceutical composition of etanercept, to a method of preparing the composition, to a kit including the composition, to a package including the composition, to a method of manufacturing the package, and to methods of treatment in which the composition and / or the container is used.

Background

Treatment of tumor necrosis factor alpha (TNF-a) -related autoimmune diseases, such as rheumatoid arthritis (including juvenile rheumatoid arthritis), psoriatic arthritis, plaque psoriasis, and ankylosing spondylitis, has been achieved by using from FDA approved drugs such as Etanercept (ENBREL®, Amgen, Wyeth) - CAS Registry Number: 185243-69-0; CAS name: Tumor necrosis factor receptor (human) 1-235 fusion protein with 236-467-immunoglobulin G1 (Fc fragment of human g1 chain)). In particular, Etanercept is a recombinant fusion protein containing soluble human TNF receptor p75 bound to the Fc portion of human immunoglobulin G1, which dimerizes through cysteine residues in the Fc fragment to form a structure of immunoglobulin type. Its preparation and uses have been previously described in the literature, including WO91 / 03553 and WO 9406476 and US 5605690 (all from Immunex), while its clinical use is discussed in B. Goffe, JC Cather, J. Am. Acad. Dermatol. 49, S105-S111 (2003); S. Nanda, J. M. Bathon, Expert Opin. Pharmacother. 5, 1175-1186 (2004). The TNFa inhibitory properties of etanercept are believed to be responsible for its clinical effects.

Initially, due to storage stability issues, etanercept (Enbrel®) was provided to clinicians and / or patients in all-purpose vials as a preservative-free, white, sterile lyophilized powder for reconstitution with 1 ml of sterile water. For injection provided, containing 0.9% benzyl alcohol, prior to subcutaneous injection. The reconstituted solutions had a pH of approximately 7.4 and were clear and colorless. Each vial contained 25 mg of etanercept, 40 mg of mannitol, 10 mg of sucrose, and 1.2 mg of tromethamine.

However, due to the drawbacks of reconstitution for clinicians and end users alike, more stable liquid formulations were developed (see WO03 / 072060 - Immunex), which could be stored for a period of time in single-use pre-filled syringes. , which could be used directly for the administration of etanercept by subcutaneous injection. Said liquid formulations of etanercept (Enbrel®) comprise a 50 mg / ml aqueous solution of etanercept, 1% w / v sucrose, 100 mM sodium chloride, 25 mM sodium phosphate, 25 mM L-arginine hydrochloride, which it has a pH of approximately 6.3. As explained in WO03 / 072060, the presence of L-arginine was considered essential for the stability and viability of the etanercept fusion protein, in particular to prevent aggregation. WO03 / 072060 describes various assays, including Size Exclusion Chromatography (SEC), Hydrophobic Interaction Chromatography (HIC), polyacrylamide gel electrophoresis and dodecyl sulfate. sodium (SDS-PAGE), to illustrate the efficacy of L-arginine as an antiplatelet for etanercept.

The difficulty of obtaining viable and stable alternative liquid formulations of etanercept has attracted the interest of a variety of researchers. For example, more recent publications such as WO2011141926 (Intas) describe alternative liquid formulations of etanercept that employ a complex range of excipients, including surfactants and chelators, among others. WO2012165914 (LG Life Sciences) describes alternative liquid formulations of etanercept in which some sulfur-containing amino acids were considered as possible stabilizers of etanercept. WO2013006454 (Biogen) describes another alternative liquid formulation of etanercept in which L-arginine is replaced by a high salt content.

Despite a significant amount of research activity in this area, it remains very difficult to achieve stable and viable arginine-free formulations, undoubtedly due to the delicate interplay of factors that affect the stability of such formulations. However, it is desirable to avoid L-arginine, especially in injectable formulations, in view of its well-known side effects, including anaphylaxis, hyperchloremic metabolic acidosis, cerebral edema, hyperkalemia, incompatibilities with certain other classes of drugs (eg, inhibitors phosphodiesterase such as sildenafil / Viagra®), swelling, diarrhea, hematuria, hives, hormonal changes, increased blood urea nitrogen, serum creatine and serum creatinine, increased inflammatory response (for example, in people with asthma or cystic fibrosis), restless legs, low back pain, nausea, night sweats and hot flashes (for example, with arginine withdrawal), numbness (especially with injections containing arginine), rash, reduced hematocrit, severe tissue damage (especially with injections containing arginine), stomach and intestinal upset, systemic acidosis and venous irritation.

Thus, there is a need for an alternative or improved liquid formulation of etanercept. Desirably, any new formulation would solve at least one of the aforementioned problems and / or at least one problem inherent in the prior art, and can adequately solve two or more of said problems. Desirably, any new formulation would avoid or minimize the occurrence of other problems. Desirably, the prior art problem (s) can be solved while reducing the complexity of the formulation.

Compendium of the invention

According to a first aspect of the present invention there is provided a liquid pharmaceutical composition comprising etanercept (suitably including any biosimilar thereof); a buffer system (or buffer agent); a tonicity regulator; and a sugar stabilizer. The composition suitably comprises (or excludes) one or more additional components defined herein in relation to a liquid pharmaceutical composition (including, for example, one or more particular amino acids; excluding arginine, etc.), optionally in any amount , concentration or form stipulated herein. The composition suitably exhibits one or more parameters or properties indicated herein in relation to a liquid pharmaceutical composition (eg, pH, osmolality, aggregation, fragmentation, protein unfolding, turbidity, etc.).

According to a second aspect of the present invention there is provided a container (for example, a pre-filled syringe, pen, intravenous bag, or a container / container containing any of the above) comprising a liquid pharmaceutical composition as defined herein. memory.

According to a third aspect of the present invention there is provided a drug delivery device (eg, syringe or pre-filled pen, or intravenous bag) comprising a liquid pharmaceutical composition as defined herein.

According to a fourth aspect of the present invention there is provided a kit of parts comprising a drug delivery device, a liquid pharmaceutical composition (or two or more parts thereof which, when reconstituted, together produce the liquid pharmaceutical composition) such as defined herein (optionally contained in a container or container) and optionally a set of instructions with directions for administration (eg, subcutaneous) of the liquid pharmaceutical composition.

According to a fifth aspect of the present invention there is provided a method of preparing a liquid pharmaceutical composition, the method comprising mixing together etanercept (suitably including any biosimilar thereof); a buffer system (or buffer agent); a tonicity regulator; and a sugar stabilizer; and optionally one or more additional components defined herein in relation to a liquid pharmaceutical composition, optionally in any stipulated amount, concentration or form; and optionally adjusting one or more parameters given herein in relation to a liquid pharmaceutical composition (eg, pH, osmolality).

According to a sixth aspect of the present invention there is provided a liquid pharmaceutical composition obtainable, obtained through or directly obtained by a method of preparing a liquid pharmaceutical composition as defined herein.

According to a seventh aspect of the present invention there is provided a method of manufacturing a drug delivery container or device, the method comprising incorporating a liquid pharmaceutical composition as defined herein into a drug delivery container or device. .

According to an eighth aspect of the present invention there is provided a container or a drug delivery device obtainable, obtained through or directly obtained by a method of manufacturing a container or a drug delivery device. as defined herein.

According to a ninth aspect of the present invention there is provided a method of treating a medical disease or disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a liquid pharmaceutical composition as defined herein. memory.

According to a tenth aspect of the present invention there is provided a liquid pharmaceutical composition as defined herein for use in therapy.

According to an eleventh aspect of the present invention there is provided the use of a liquid pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a disease or disorder.

According to a twelfth aspect of the present invention there is provided a method for treating an autoimmune disease related to tumor necrosis factor alpha (TNF-a) in a patient in need of said treatment, said method comprising administering to said patient a therapeutically effective amount of a liquid pharmaceutical composition as defined herein.

According to a thirteenth aspect of the present invention there is provided a liquid pharmaceutical composition as defined herein for use in treating an autoimmune disease related to tumor necrosis factor alpha (TNF-a).

According to a fourteenth aspect of the present invention there is provided the use of a liquid pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of an autoimmune disease related to tumor necrosis factor alpha (TNF-a ).

According to a fifteenth aspect of the present invention there is provided a method for treating rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, plaque psoriasis and / or ankylosing spondylitis in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a liquid pharmaceutical composition as defined herein.

According to a sixteenth aspect of the present invention there is provided a liquid pharmaceutical composition as defined herein for use in treating rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, plaque psoriasis, and / or ankylosing spondylitis.

According to a seventeenth aspect of the present invention there is provided the use of a liquid pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, psoriasis. plaque and / or ankylosing spondylitis.

In further aspects, the invention provides a liquid pharmaceutical composition, a container, a drug delivery device, a kit of parts, a method of preparing a liquid pharmaceutical composition, a method of manufacturing a container, or a drug delivery device. drugs, a method of treatment, a liquid pharmaceutical composition for use, and a use of a liquid pharmaceutical composition in the manufacture of a drug, essentially as defined herein (including in any of the twenty aspects mentioned above) except that, rather than being specific for "etanercept" (and biosimilars thereof), the invention can be applied (and therefore defined as related) to any TNF-α inhibitor protein, including a fusion protein, antibody ( anti-TNF-a) antibody, antibody fragment or any biosimilar thereof, although fully suitably a fusion protein that inhibits e activity of human TNF-a. Suitably, the fusion protein is a therapeutically effective drug (at least when administered in adequate amounts to a patient in need) (or a biosimilar thereof; see below for definitions of biosimilars in relation to etanercept, which equally applies to all TNFa inhibitory proteins), suitably one that has received FDA approval. As such, any reference herein to "etanercept" may be construed, unless inconsistent with it, as a reference to any TNFa inhibitor protein for the purpose of these additional aspects of the invention (regardless of whether this is refers to absolute or relative quantities, concentrations, parameters or properties, or regardless of whether this refers to certain definitions, such as what a biosimilar consists of).

One of these additional aspects of the present invention provides a liquid pharmaceutical composition comprising a TNFa inhibitor protein (suitably including any biosimilar thereof); a buffer system (or buffer agent); a tonicity regulator; and a sugar stabilizer; the composition optionally comprising (or excluding) one or more additional components defined herein in relation to a liquid pharmaceutical composition (eg, including one or more particular amino acids; excluding arginine, etc.), optionally in any amount, concentration or form stipulated herein; and the composition optionally exhibiting one or more parameters or properties indicated herein in relation to a liquid pharmaceutical composition (eg, pH, osmolality, aggregation, fragmentation, protein unfolding, turbidity, etc.).

Suitably, the TNFa inhibitory protein treats inflammation. Suitably, TNFa inhibitor protein can be used to treat rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, plaque psoriasis, ankylosing spondylitis, inflammatory bowel disease, psoriasis, hidradenitis suppurativa, and asthma. refractory. In a particular embodiment, the TNFa inhibitor protein is selected from a fusion protein and a monoclonal antibody. In a particular embodiment, the fusion protein is etanercept. In a particular embodiment, the monoclonal antibody is selected from the group consisting of infliximab, adalimumab and certolizumab pegol.

Any features, including optional, suitable and preferred features, described in connection with any particular aspect of the invention may also be features, including optional, suitable and preferred features, of any other aspect of the present invention.

Detailed description of the invention

Definitions

Unless otherwise indicated, the following terms used in the specification and claims have the following meanings presented below.

Throughout the description and claims of this specification, the words "comprise" and "contains" and their variations mean "including, but not limited to", and are not intended to exclude (or exclude) other moieties, additives , components, integers or stages. Throughout the description and claims of this specification, the singular encompasses the plural unless the context requires otherwise. In particular, when using the indeterminate article, it is to be understood that the specification contemplates both plurality and singularity, unless the context requires otherwise.

The particularities, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood as applicable to any other aspect, embodiment or example described herein unless incompatible with the same. All the features described in this specification (including any of the appended claims, the abstract and the drawings) and / or all the steps of any method or process thus disclosed, can be combined in any combination, except combinations in which at least some of said characteristics and / or stages are mutually exclusive. The invention is not limited to the details of any of the above embodiments. The invention extends to any new feature, or to any new combination of the features described in this specification (including any of the appended claims, the abstract, and the drawings), or to any new step, or new combination of the steps of any method. or process thus revealed.

The reader's attention is directed to all articles and documents that are filed at the same time or prior to this specification in connection with this application and that are open to public inspection with this specification, and the content of all such articles and documents are incorporated herein by reference.

For the avoidance of doubt, it is hereby stated that the information described above in this specification under the heading "Background" is relevant to the invention and should be read as part of the description of the invention.

Suitably, unless otherwise stated, when referring to a parameter (e.g. pH, pKa, etc.) or a state of a material (e.g. liquid, gas, etc.) that may depend on the pressure and / or temperature, suitably in the absence of further clarification, said reference is related to said parameter at standard ambient pressure and temperature (SATP). SATP consists of a temperature of 25 ° C (298.15 K, 77 ° F) and an absolute pressure of 100 kPa (14.504 psi, 0.987 atm).

Unless otherwise indicated, any reference herein to an "average" value is intended to be related to the average value.

References herein to "etanercept" include the commercially available drug as Enbrel®, as well as etanercept as defined in WO91 / 03553 and WO 9406476 and US 5605690 (all from Immunex) and elsewhere in the art. , and also biosimilars thereof. Etanercept is a homodimeric fusion protein formed from the combination of two soluble human 75 kilodalton TNF receptors, each bound to an Fc portion of an IgG1. Etanercept is suitably produced by recombinant DNA. This suitably fuses the TNF receptor with the Fc portion of the IgG1 antibody. Relevant recombinant DNA can be produced by isolating the DNA sequence encoding the human gene for soluble TNF receptor 2 (ie, a receptor that binds TNFα); and also the DNA sequence encoding the human gene for the Fc portion of IgG 1; before joining these two DNA sequences together. The joined DNA sequence is then expressed to produce a protein containing the protein sequence for the soluble TNF receptor 2 fragment bound to the IgG 1 Fc protein. The sequence listing for the monomer component of etanercept and a method to form a crystalline form thereof are also described in US7276477 (identified therein as SEQ ID No: 4). The dimers of these fusion proteins are formed when monomers are held together by three disulfide bonds that form between the immunoglobulin portions of the two monomers.

References herein to "etanercept" may include biosimilars that, for example, may share at least 75%, suitably at least 80%, suitably at least 85%, suitably at least 90%, suitably at least 95%, suitably at least 96%, suitably at least 97%, suitably at least 98% or totally suitably at least 99% protein sequence identity with any of the protein sequences described in the technique related to etanercept. Alternatively or additionally, references herein to "etanercept" may include biosimilars exhibiting at least 75%, suitably at least 80%, suitably at least 85%, suitably at least 90%, suitably at least a 95%, suitably at least 96%, suitably at least 97%, suitably at least 98% or totally suitably at least 99% protein sequence homology to any of the protein sequences described in the art related to etanercept. Alternatively or additionally, a biosimilar may have a (slightly) different glycosylation profile, even if the protein sequence is substantially the same or different to the extent specified above.

Suitably herein, the etanercept employed in the compositions and methods of the invention is "substantially purified," which suitably means that the etanercept is a polypeptide that is substantially free of other polypeptides present in the environment in which it is presented. naturally or in which it has been produced; a preparation of a polypeptide that has been substantially purified contains at least 90% by weight (or at least 95%, at least 98% or at least 99% by weight) of that polypeptide, wherein the weight of the polypeptide includes any carbohydrate, lipid, or other residues covalently attached to the polypeptide. A substantially purified polypeptide preparation may contain variation between polypeptide molecules within of preparation, with respect to the degree and type of glycosylation or other post-translational modification, or with respect to conformation or degree of multimerization.

The term "biosimilar" (also known as follow-on biologics) is well known in the art, and one of ordinary skill in the art would readily appreciate when a drug substance would be considered a biosimilar of etanercept. Furthermore, such "biosimilars" would have to be officially approved as "biosimilars" for commercialization before said "biosimilar" is sold on the open market. The term "biosimilar" is generally used to describe later versions (usually from a different source) of "innovative biopharmaceuticals" ("biologics" whose drug substance is produced by a living organism or derived from a living organism or by DNA methodologies recombinant or controlled gene expression) that have previously obtained an official marketing authorization. Since biologics have a high degree of molecular complexity and are generally sensitive to changes in manufacturing processes (for example, if different cell lines are used in their production), and since follow-on manufacturers generally They do not have access to the creator's molecular clone, cell bank, knowledge of the production process, or the active principle itself (only the pharmaceutical product marketed by the innovator), any "biosimilar" may not be exactly the same as the pharmaceutical product innovative.

For the purposes of various molar calculations (for example, for the molar ratios between etanercept and another component of the liquid pharmaceutical composition of the invention), the molecular weight of etanercept can be assumed to be 51,234.90 g / mol (molecular weight reference) based on the details described in the CAS database for CAS Registry Number: 185243-69-0, Etanercept, where the molecular formula is assumed to be C ²²²⁴ H ³⁴⁷⁵ N ⁶²¹ O ⁶⁹⁸ S ³⁶ . As such, a liquid pharmaceutical composition containing 50 mg / ml of etanercept can be considered a 0.976 mM (or 976 pM) solution of etanercept. This is not intended to be limiting in any way with respect to the nature of the etanercept biosimilars covered by the scope of the present invention, nor the level of glycosylation, either of which can affect actual molecular weight. However, when a biosimilar has a different molecular weight, the above-mentioned reference molecular weight has to be suitably used in order to assess whether or not said biosimilar falls within the scope of any molar definition stipulated within this specification. Therefore, the number of moles in a known weight of said biosimilar has to be calculated, only for the purposes of this invention, using the reference molecular weight indicated above.

As used herein, the terms "buffer" or "buffer solution" refer to a generally aqueous solution comprising a mixture of an acid (usually a weak acid, eg, phosphoric acid or one or more species of hydrogen phosphate) and its conjugate base (eg a phosphate, eg sodium phosphate or derivatives thereof) or alternatively a mixture of a base (usually a weak base, eg histidine) and its conjugated acid (eg protonated histidine salt ). The pH of a "buffer solution" will change only slightly with the addition of a small amount of strong acid or base due to the "buffer effect" imparted by the "buffer agent".

As used herein, a "buffer system" comprises one or more buffering agents and / or acid / base conjugate (s) thereof, and more suitably comprising one or more buffering agents and acid / base conjugate (s) thereof. . Unless otherwise indicated, any concentration stated herein in relation to a "buffer system" (ie, a buffer concentration) appropriately refers to the combined concentration of the buffer agent (s) and / or the buffer (s). the acid / base conjugates thereof. In other words, the concentrations stated here in relation to a "buffer system" appropriately refer to the combined concentration of all relevant buffer species (ie species in dynamic equilibrium with each other, for example phosphate (s) / acid ( s) phosphoric (s)). As such, a given concentration of a phosphate and / or histidine buffer system generally refers to the combined concentration of histidine and the imidazolium form of histidine and / or phosphate (s) and / or phosphoric acid (s). ). In the case of phosphate buffer systems, these concentrations are usually easy to calculate with reference to the input amounts of phosphate salts (including any hydrogen phosphate salts, for example monohydrogen phosphate, dihydrogen phosphate and / or trihydrogen phosphate). In the case of histidine, these concentrations are usually easy to calculate with reference to the input amounts of histidine or a salt thereof. The total pH of the composition comprising the relevant buffer system is generally a reflection of the equilibrium concentration of each of the relevant buffer species (i.e., the equilibrium between the buffer agent (s) and the acid / base conjugate (s) thereof).

As used herein, the term "buffering agent" refers to an acid or base component (usually a weak acid or a weak base) of a buffer or a buffer solution. A buffering agent helps to maintain the pH of a given solution at or near a predetermined value, and buffering agents are generally chosen to complement the predetermined value. A buffering agent is suitably a single compound that gives rise to a desired buffering effect, especially when said buffering agent is mixed with (and is suitably capable of exchanging protons with) an appropriate amount (depending on the predetermined desired pH) of its "conjugate of Corresponding "acid / base", or if the required amount of its corresponding "acid / base conjugate" is formed in situ - this can be achieved by adding strong acid or base until the required pH is reached. As an example:

• A phosphate "buffer" is suitably a phosphate salt, for example a sodium phosphate (which may include one or a mixture of two or more phosphates, such as a mixture of monosodium phosphate, monobasic disodium phosphate and / or trisodium phosphate ) properly mixed with its acid / base conjugate, phosphoric acid. Such a buffer system can be formed simply by mixing a certain amount of phosphate salt (s) with a specified amount of phosphoric acid. Alternatively, however, such a buffer can be formed by adding a given amount of a base, suitably a strong base (e.g. sodium hydroxide) to phosphoric acid until the desired pH (and therefore the desired balance is reached sodium acetate / acetic acid). Herein, unless otherwise indicated, any given concentration relative to a phosphate buffer or phosphate buffer is suitably referred to as the combined concentration of the buffer agent (s) (eg, sodium phosphate) and / or acid / base conjugate (s) thereof (eg phosphoric acid). The person skilled in the art will be able to easily calculate these concentrations. Such concentrations can be calculated with reference to the combined concentrations of buffer agent (s) and acid / base conjugate (s), when a buffer system is formed simply by mixing buffer agent (s) and acid / base conjugate (s) together. base. Alternatively, when a buffer system is formed by mixing the buffer agent (s) or acid / base conjugate (s) with a pH adjuster (eg, strong acid or strong base) to produce a mixture of each, these concentrations can be properly calculate with reference to the initial amounts / concentrations of the buffer agent (s) or the acid / base conjugate (s), respectively. For example, when a buffer system is formed using a known amount / concentration of phosphoric acid that is mixed with a pH adjuster (e.g., sodium hydroxide) until the desired pH is reached, the concentration of the buffer system can be calculated. with reference to the initial amount of phosphoric acid.

• A citrate "buffering agent" is suitably a salt of citrate, for example sodium citrate, suitably mixed with its acid / base conjugate, citric acid. Such a "buffer system" can be formed simply by mixing a given amount of sodium citrate with a given amount of citrate acid. Alternatively, however, such a buffer can be formed by adding a given amount of a base, suitably a strong base (e.g. sodium hydroxide) to citric acid until the desired pH (and therefore the desired balance is reached sodium citrate / citric acid). Herein, unless otherwise indicated, any given concentration in relation to a citrate buffer or a citrate buffering agent suitably refers to the combined concentration of the buffer agent (s) (eg, sodium citrate) and / or the acid / base conjugates thereof (eg citric acid). The person skilled in the art will be able to easily calculate said concentration. Such concentrations can be calculated with reference to the combined concentrations of buffer agent (s) and acid / base conjugate (s), when a buffer system is formed simply by mixing buffer agent (s) and acid / base conjugate (s) together. base. Alternatively, when a buffer system is formed by mixing the buffer agent (s) or acid / base conjugate (s) with a pH adjuster (eg, strong acid or strong base) to produce a mixture of each, these concentrations can be properly calculate with reference to the initial amounts / concentrations of the buffer agent (s) or the acid / base conjugate (s), respectively. For example, when a buffer system is formed using a known amount / concentration of citric acid that is mixed with a pH adjuster (e.g. sodium hydroxide) until the desired pH is reached, the concentration of the buffer system can be calculated with reference to the initial amount of citric acid.

As used herein, an "acid / base conjugate" refers to the conjugated acid or the conjugated base (whichever is relevant at a particular pH, usually the conjugated acid in the context of the present invention) of a "buffer agent" particular. The acid / base conjugate of a phosphate buffer (e.g. sodium phosphate) is suitably phosphoric acid (although this may suitably include higher hydrogen phosphate salts, such as mono- or di-hydrogen phosphates, which are potentially acidic conjugates. with phosphate, or lower hydrogen phosphate salts thereof). The acid / base conjugate of a citrate buffer (eg, sodium citrate) is suitably citric acid.

As used herein, the term "buffer species" refers to the particular species (excluding any associated counter anions or counter cations, that is, ignoring sodium counter ions for phosphate (s) / phosphoric acid (s) systems ), and ignoring the chloride or hydroxide counter ions for the histidine / imidazolium-histidine systems) of a given buffer system that are in dynamic equilibrium (and in proton exchange) with each other. For example, the anions of phosphate, monohydrogen phosphate and dihydrogen phosphate and phosphoric acid may together constitute "phosphate buffer species" of a "phosphate buffer system" (although, depending on the pH, some may prevail more than others within the system tampon). For example, citrate anions and citric acid together constitute a "citrate buffer species" of a "citrate buffer system".

Since it is often somewhat difficult to define the amounts (either absolute or relative) of a buffer system with reference to weight (since the total weight will depend on the desired pH, which will influence the amount of counter ions present), herein Weight-based amounts can instead be determined with reference to a theoretical weight of an arbitrary or relevant "buffer species." Generally, in any given set of "buffer species" at least two species are present (in relative amounts that can only be determined with reference to pH), each with a different molecular weight (which generally differs only by 1, although for phosphate buffers multiple phosphate species may be present). Therefore, to allow for viable weight calculations and references, for the purposes of this specification, the weight of any given set of "buffer species" is given as a theoretical weight based on only one of the buffer species. Depending on the buffer system in question, this may suitably be the most acidic of the buffer species (i.e., the most protonated at any given pH) or the most basic of the buffer species (ie, the least protonated at any given pH), or it may simply be one of the buffer species. By way of example, in a phosphate system, the phosphate buffer species may consist of phosphate anions (ignoring countercations) and phosphoric acid (and / or one or more corresponding hydrogen phosphates). Therefore, the weight of the "buffer species" can be calculated as if phosphoric acid were the only species present in the buffer system (although there are clearly phosphate species present along with phosphoric acid, and even if it is only actually present a minimum of phosphoric acid with the given pH). Thus, any reference to a weight or a weight ratio that implies a "phosphate buffer species" is appropriately related to the theoretical weight of phosphoric acid equivalents within the buffer system. As such, when a composition is formed by adding a pH adjuster (eg, sodium hydroxide) to a fixed amount of phosphoric acid, the original weight of phosphoric acid can be considered as the weight of the "buffer species" regardless of the pH. final. Alternatively, if the concentration (i.e. molarity) of a buffer system is known (or if the relevant buffer system is formed by adding an acidic pH adjustment, such as phosphoric acid, to a fixed amount of phosphate species), it is can be converted to a "buffer species" weight with reference to the molecular weight of the most acidic form of the relevant buffer species (eg, phosphoric acid), and ignoring the fact that phosphate anions are also present. A similar principle is applicable to citric acid for citrate buffer systems.

Those skilled in the art will understand that many buffer species can take a number of different forms. Furthermore, acids, such as phosphoric acid, may themselves take various forms - collectively "phosphoric acids" - and the present invention suitably includes any of these forms.

Unless otherwise indicated, references herein to an "amino acid" or to "amino acids", whether specific (eg, arginine, histidine) or general (eg, any amino acid), in the context of their presence or otherwise within the compositions (especially liquid pharmaceutical compositions of the invention) are related to the corresponding free amino acid (s) (regardless of their protonation state and / or salt form, although to maintain consistency, the amounts are properly calculated with reference to the free amino acid per se). This can suitably include natural and / or artificial amino acids. Unless otherwise indicated, such references are not intended to relate to amino acid residue (s) covalently incorporated as part of a larger compound (as opposed to a composition comprising multiple compounds), such as a peptide. or a protein (where said amino acid residues are linked through peptide bonds). As such, although etanercept, as a protein, contains amino acid residues, it is not considered to comprise any "free amino acids". By way of example, a composition defined as "arginine free" does not contain any free arginine, but can still include one or more proteins (eg, etanercept) that do comprise arginine residues.

Unless otherwise indicated, references herein to one or more "amino acids", whether specific or general, are suitably related to the L-stereoisomers or at least to a mixture comprising the L-stereoisomers. suitably, said "amino acids" are L-amino acids.

The term "substantially free", when used in relation to a given component of a composition (eg, "a liquid pharmaceutical composition substantially free of arginine"), refers to a composition to which essentially none of the components have been added. said components. As explained above, such references have no relation to the presence of amino acid residue (s) within a protein structure. When a composition is "substantially free" of a given component, said composition suitably comprises no more than 1.0% by weight of said component, suitably no more than 0.1% by weight of said component, suitably no more than 0.01% by weight of said component, suitably no more than 0.001% by weight of said component, suitably no more than 0.0001% by weight of said component, suitably no more than 0.00001% by weight suitably no more than 0.000001% by weight, suitably no more than 0.0000001% by weight thereof, fully suitably no more than 0.0001 parts per billion (by weight).

The term "completely free", when used in relation to a given component of a composition (eg, "a liquid pharmaceutical composition substantially free of arginine"), refers to a composition that does not contain any such component. As explained above, such references do not relate to the presence of amino acid residue (s) within a protein structure.

Herein, in the context of the present specification, a "strong acid" is suitably one that has a pK ^a of -1.0 or less, while a "weak acid" is suitably one that has a pK ^a of. 2.0 or more. Herein, in the context of the present specification, a "strong base" is suitably one whose conjugated acid has a pK ^a of 12 or more (suitably 14 or more), while a "weak base" is suitably one whose Conjugated acid has a pK ^a of 10 or less.

Herein, a "stabilizer" refers to a component that facilitates the maintenance of the structural integrity of the biopharmaceutical drug, particularly during freezing and / or lyophilization and / or storage (especially when exposed to stress). This stabilizing effect can arise for a variety of reasons, although normally such stabilizers can act as osmolytes that mitigate protein denaturation. Typical stabilizers include sugar stabilizers, such as a sugar polyol (for example, mannitol, sorbitol) and / or a disaccharide (e.g. trehalose, sucrose, maltose, lactose), although the liquid pharmaceutical compositions of the invention include a stabilizer, at least one of them being a sugar stabilizer (i.e. a polyol sugar or a disaccharide).

As used herein, a "tonicity modifier" or "tonicity regulator" refers to a reagent whose inclusion within a composition adequately contributes to (or increases) the overall osmolality and osmolarity of the composition. Suitably, a tonicity regulator, as used herein, includes an agent whose function is to make a solution similar in osmotic characteristics to physiological fluids.

Herein, references to specific amounts of a given component of a composition, especially a buffer agent (or buffer system), stabilizer, amino acid, surfactant, or tonicity regulator, are suitably related to amounts of the pure anhydrous form of the relevant component (or compositions formed by using said amounts of the pure anhydrous form), although said component may be used in a non-anhydrous form when forming the composition. The amounts of any corresponding non-anhydrous form (eg monohydrates, dihydrates, etc.) can be easily calculated by simply using the appropriate multiplier.

Herein, references to any given component of a composition, unless otherwise indicated, suitably include alternative forms of said component or include said component when formed from alternative forms, such as any salt, free base, free acid, solvate or complex thereof, especially with respect to a buffer agent (or buffer system), stabilizer, amino acid, surfactant or tonicity regulator. As such, reference to a composition comprising a given compound (eg arginine) includes any equivalent composition comprising said component in an alternative form (eg arginine hydrochloride), or any equivalent composition formed by incorporating a alternative form of arginine (for example, arginine hydrochloride). However, unless otherwise stated, any stipulated amount with respect to the given component (especially when given in terms of weight or concentration / weight ratio) will appropriately refer to the stipulated amount of that component, not to the amount of any alternative form thereof (eg salt). It will be necessary to apply a conversion factor to establish the equivalent amount of said component in any alternative form, such as salt, free base, free acid, solvate or complex thereof; this can be calculated by the person skilled in the art, simply with reference to relevant molecular weights of the relevant species. As such, wherever the invention is defined with reference to a stipulated amount of "arginine", this covers compositions containing that amount of arginine in hybrid ionic form, even if the source of arginine is an alternate form, and even if arginine exists within the composition in an alternative form (eg, which may well be pH dependent).

As used herein, the term "pharmaceutical composition" refers to a formulation of a pharmaceutical active ingredient that renders the biological activity of the active ingredient therapeutically effective, but does not include other ingredients that are obviously toxic to a subject to whom it is used. intended to administer the formulation.

As used herein, the term "stable" refers generally to the physical stability and / or the chemical stability and / or the biological stability of a component, usually a pharmaceutical active ingredient or a composition thereof, during preservation / storage. .

It should be noted that references to "treat" or "treatment" include both prophylaxis and mitigation of established symptoms of a condition. Therefore, "treating" or "treating" a condition, disorder, or condition includes: (1) preventing or delaying the onset of clinical symptoms of the condition, disorder, or condition that develops in a human that may be affected by the state, disorder or condition or predisposed to it, but not yet experiencing or displaying clinical or subclinical symptoms of the condition, disorder or condition, (2) inhibiting the condition, disorder or condition, that is, arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one of its clinical or subclinical symptoms, or (3) alleviate or attenuate the disease, that is, cause the regression of the state, disorder or condition or of at least one of its clinical or subclinical symptoms.

In the context of the present invention, a "therapeutically effective amount" or "effective amount" of etanercept means an amount that is effective when administered to a mammal to treat a disease or disorder, in the prophylactic and therapeutic aspect, and the antibody it is effective in treating the diseases in question.

The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the mammal to be treated.

The term "human TNF-a" refers to the human cytokine that exists in a secreted form of 17 kD and a membrane-associated form of 26 kD, and in a biologically active form, TNF-a could be observed as a trimer of a 17 kD molecule covalently attached. Its specific structure can be found in Pennica, D. et al. (1984) Nature 312: 724-729; Davis, J. M. et al. (1987) Biochemistry 26, 1322-1326; and Jones, E. Y. et al. (1989) Nature 338: 225-228.

Herein, the amounts stated for components and ingredients, regardless of whether they are specified in terms of "parts", ppm (parts per million), percentages (%, for example% by weight) or proportions, they are intended as amounts by weight, unless otherwise indicated.

When the amount or concentration of a particular component of a given composition is specified as a percentage by weight (% by weight or% w / w), said percentage by weight refers to the percentage of said component by weight with respect to the total weight of the composition as a whole. Those skilled in the art will understand that the sum of the weight percentages of all components of a composition (whether or not specified) will total 100% by weight. However, when not all components are listed (for example, when the compositions are said to "comprise" one or more particular components), the remaining weight percent may optionally be up to 100% by weight of unspecified ingredients ( for example a diluent, such as water, or other non-essential but suitable additives).

As used herein, unless otherwise indicated, the term "parts" (eg parts by weight, pep), when used in relation to multiple ingredients / components, refers to relative relationships between such multiple ingredients / components. . Expressing molar or weight ratios of two, three, or more components gives the same effect (for example, a molar ratio of x, y, z is ^{xi: yi: zi} respectively, or an interval ^{X1-X2: yi - y 2 : Z1-Z2} ). Although in many embodiments the amounts of individual components within a composition may be indicated as a "wt%" value, in alternate embodiments any or all of these wt% values may be converted to parts by weight (or proportions relative) to define a multi-component composition. This is so because the relative proportions between the components are often more important than the absolute concentrations of the same in the liquid pharmaceutical compositions of the invention. When a composition comprising multiple ingredients is described only in terms of parts by weight (i.e. to indicate only relative proportions of ingredients), it is not necessary to stipulate the absolute amounts or concentrations of those ingredients (either in total or individually), since the advantages of the invention may be derived from the relative proportions of the respective ingredients rather than their absolute amounts or concentrations. However, in certain embodiments, such compositions consist or consist essentially of the stated ingredients and a diluent (eg, water).

When a composition is said to comprise a plurality of stipulated ingredients (optionally in stipulated amounts of concentrations), said composition may optionally include additional ingredients other than those stipulated. However, in certain embodiments, when a composition is said to comprise a plurality of stipulated ingredients, it may in fact consist or consist essentially of all of the stipulated ingredients.

Herein, when a composition is said to "consist essentially of" a particular component, said composition suitably comprises at least 85% by weight of said component, suitably at least 90% by weight thereof, suitably at least one 95% by weight thereof, fully suitably at least 99% by weight thereof. Suitably, when a composition is said to "consist essentially of" a particular component, it consists of that component except for one or more trace impurities.

As used herein, the term "particle size" or "pore size" refers respectively to the length of the longest dimension of a given particle or pore. Both sizes can be measured using a laser particle size analyzer and / or electron microscopes (eg, tunneling electron microscope, TEM, or scanning electron microscope, SEM). The particle count (for any given size) can be obtained using the protocols and equipment described in the Examples, which refer to the particle count of the sub-visible particles.

Herein, unless otherwise indicated, all chemical nomenclature may be defined in accordance with IUPAC definitions.

Liquid pharmaceutical composition

The present invention provides a liquid pharmaceutical composition, suitably as defined herein. The composition suitably comprises a protein, suitably one that inhibits human TNF-α activity, suitably to prevent it from activating TNF receptors. Suitably, the liquid pharmaceutical composition comprises a fusion protein, suitably a fusion protein comprising a TNF receptor bound to an Fc portion of an IgG1. Fully suitably, the pharmaceutical composition comprises etanercept, including any biosimilars thereof.

The composition suitably comprises a buffer system or a buffer agent. The composition suitably comprises a buffer system, and the pH of the composition is suitably between pH 5.7 and 6.9. Suitably the buffer system maintains the composition at a (substantially) constant pH (suitably varying by no more than / - 0.5 pH units, suitably no more than / - 0.2 pH units, suitably no more than / - 0.1 pH units under standard conditions of storage and use).

The composition suitably comprises a tonicity regulator. Suitably, the tonicity regulator serves to provide the composition with osmolality or tonicity close to or (substantially) isotonic with body fluids.

The composition suitably comprises a stabilizer, suitably a sugar stabilizer.

Suitably the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, or in a weight ratio of arginine to etanercept of at most 1: 3,000 (i.e. less or equal to one part by weight of arginine for every 3,000 parts by weight of etanercept). Alternatively or additionally, the composition may suitably include one or more additional components defined herein in relation to a liquid pharmaceutical composition (eg, including particular amino acid (s), excluding particular amino acid (s)). , optionally in any amount, concentration, or form stipulated herein; and wherein the composition optionally exhibits one or more parameters or properties indicated herein in relation to a liquid pharmaceutical composition (eg, pH).

Advantageously, the present invention provides alternative and improved liquid pharmaceutical compositions, which generally exhibit better stability and workability than those of the prior art. As illustrated herein (see examples), the liquid pharmaceutical formulations of the present invention have comparable or even improved characteristics compared to conventional etanercept formulations, for example the commercially available formulation Enbrel®, when subjected to different stress conditions (thermal, mechanical and light). Since most preferably the compositions of the invention are (substantially) free of arginine, they offer significant benefits associated with the avoidance of arginine-specific side effects. It was considered surprising that such good stability performance can be achieved using less complex formulations, with fewer excipients and generally without arginine, in view of the general teachings of the prior art.

Etanercept

As detailed in the definitions section, references herein to "etanercept" include the parent drug substance (commercially available as Enbrel®), as well as etanercept as defined in WO91 / 03553 and Wo 9406476 and US 5605690 (all from Immunex) and elsewhere in the art, and also biosimilars thereof.

Additionally, references to "etanercept" herein may include biosimilars, as defined in the definitions section. The person skilled in the art will readily appreciate the scope of the term "etanercept" in the context of the invention.

The etanercept used in the compositions of the invention is suitably substantially purified.

Herein, the etanercept employed in the compositions and methods of the invention is suitably "substantially purified", suitably containing at least 90% by weight, more suitably at least 95% by weight, of the relevant polypeptide, including the weight of the polypeptide any carbohydrate, lipid, or other residues covalently bound to the polypeptide, despite some tolerable variation in the degree and type of glycosylation or other post-translational modification, or with respect to conformation or degree of multimerization.

The medical indications and role of etanercept are clarified above.

In one embodiment, the liquid pharmaceutical composition comprises etanercept in a concentration of from about 5 to about 150 mg / ml, suitably from about 20 to about 70 mg / ml, suitably from about 15 to 35 mg / ml, or from about 40 to 60 mg. / ml. In one embodiment, etanercept is present in a concentration of about 20 to about 30 mg / ml, quite suitably in a concentration of about 25 mg / ml. In one embodiment, etanercept is present at a concentration of about 45 to about 55 mg / ml, quite suitably at a concentration of about 50 mg / ml.

Buffer and pH systems

Suitably the liquid pharmaceutical composition is a buffered solution. The pH of the buffered solution is suitably stabilized by a buffer agent (or a buffer system), suitably in combination with an acid / base conjugate of the buffer. As such, the liquid pharmaceutical composition suitably comprises a buffering agent (or at least one buffering agent) as defined herein. Preferably, the liquid pharmaceutical composition further comprises an acid / base conjugate, wherein said acid / base conjugate corresponds to the conjugated acid or the conjugated base of the buffering agent, depending on whether the buffering agent itself is a base or an acid, respectively. . Taken together, the buffering agent and its acid / base conjugate (which may be plural in some cases, such as phosphate buffer systems comprising numerous buffer species) can be considered a "buffer system". Thus, the liquid pharmaceutical composition suitably comprises a "buffer system" (suitably comprising one or more buffering agents and one or more acid / base conjugates thereof), and any stipulated concentration in relation to the buffer system generally refers to the combined concentrations of the buffer agent (s) and any acid / base conjugate thereof. Any "buffer system" suitably comprises a weak acid and a weak base (see definitions above).

Suitably the buffer system is a phosphate buffer system and / or a citrate buffer system. Suitably, the liquid pharmaceutical composition comprises at most one buffer system, wholly suitably consisting of a phosphate buffer system or a citrate buffer system.

Suitably the buffering agent is a phosphate buffering agent. Suitably the phosphate buffering agent is a phosphate (or hydrogen phosphate salt), suitably comprising anionic phosphate species and one or more pharmaceutically acceptable countercations. A suitable phosphate salt may include a metal phosphate salt (for example, an alkali metal phosphate or an alkaline earth metal phosphate, which may include one or more of various hydrogen phosphates), or a non-metal phosphate salt (for example, ammonium phosphate, triethylammonium phosphate, which may include one or more of various hydrogen phosphates). In a particular embodiment, the buffer agent (and the phosphate salt) comprises sodium phosphate (s) (which can include sodium hydrogen phosphate and / or sodium dihydrogen phosphate, as well as optionally include trisodium phosphate).

Suitably, the liquid pharmaceutical composition comprises one or more acid / base conjugates of the buffer agent (s), wholly suitably phosphoric acid (or a hydrogen phosphate) as the conjugated acid of a phosphate salt. The combination of the buffer agent (s) and their acid / base conjugate (s) constitutes a buffer system. Suitably, the liquid pharmaceutical composition comprises the buffer agent (s) and their corresponding acid / base conjugate (s), suitably so that the buffer agent (s) and their acid conjugate (s) / base together are present at a level (ie, absolute amount or concentration) and in a relative amount (or concentration) sufficient to provide the desired pH for the composition. The buffer system can be formed in a variety of ways, especially when it comes to phosphate buffer systems, where multiple buffer species may be present. The buffer system can be formed simply by mixing the buffer agent (s) with its acid / base conjugate (s), or alternatively it can be formed by mixing an acid or base with the buffer agent (s) or its conjugate (s) ( s) of acid / base to form in situ the desired mixture of buffer agent (s) and acid / base conjugate (s). For example, the buffer system can be formed simply by mixing a phosphate buffer (e.g., sodium phosphate and / or related hydrogen phosphate (s)) with its acid / base conjugate (i.e., phosphoric acid) or indeed with a strong acid (eg hydrochloric acid), suitably in an appropriate ratio to provide the desired pH. Alternatively, the buffer system can be formed by adding a base (e.g., sodium hydroxide) to the acid / base conjugate (i.e., related phosphoric acid and / or hydrogen phosphate (s)) of the phosphate buffer, suitably in a appropriate amount to provide the desired pH and mixture of buffering agent (eg, sodium phosphate (s)) and corresponding acid / base conjugate (s) (ie, phosphoric acid). Alternatively, either of the two buffer system formation methods can be employed, and the pH can be judiciously adjusted by adding either more acid (suitably strong acid, like HCl), or more base (suitably strong base, like sodium hydroxide).

Fully suitably, the buffer system is or at least comprises a phosphate buffer system, suitably comprising a mixture of one or more phosphate salts (suitably selected from tribasic phosphate salts, dibasic hydrogen phosphate salts or dihydrogen phosphate salts. monobasic) and / or phosphoric acid.

In some embodiments, the buffer system comprises two or more buffer systems, for example a phosphate buffer system and another buffer system, such as a citrate or histidine buffer system. With respect to the phosphate buffer system, the buffer agent and the buffer species can be as described above. A citrate and / or histidine buffer system can be formed following the same or similar principles as the phosphate buffer system.

Suitably, the liquid pharmaceutical composition has a pH greater than or equal to pH 5.5, more suitably greater than or equal to pH 5.8, fully suitably greater than or equal to pH 6.2.

Suitably the liquid pharmaceutical composition has a pH less than or equal to pH 7.0, more suitably less than or equal to pH 6.8, fully suitably less than or equal to pH 6.4.

Suitably, the liquid pharmaceutical composition has a pH between 6.1 and 6.5. More suitably, the liquid pharmaceutical composition has a pH between pH 6.2 and 6.4, fully suitably a pH of 6.3 (optionally / -0.2).

Within these pH ranges, the predominant buffer species present within a phosphate buffer system are monohydrogen phosphate and dihydrogen phosphate.

Within these pH ranges, the predominant buffer species present within a citrate buffer system are citrate monohydrogen (ie citrate dianion) and citrate (ie citrate trianion).

Suitably, the liquid pharmaceutical composition comprises a buffer system (suitably a phosphate buffer system, suitably comprising a phosphate buffering agent; or a citrate buffer system, suitably comprising a citrate buffering agent) at a concentration of about 1 to 50mM, more suitably from about 10 to 40mM, fully suitably from about 20mM to 30mM. In a particular embodiment, the liquid pharmaceutical composition comprises a buffer system in a concentration of approximately 25 mM.

In one embodiment, the liquid pharmaceutical composition comprises a phosphate buffer system comprising two or more buffer species selected from phosphoric acid, dihydrogen phosphate, monohydrogen phosphate and phosphate (and / or derivatives thereof, eg o-phosphoric acid) , suitably with sodium counter ions for any phosphate species, in a collective concentration of about 1 to 50 mM, suitably of about 10 to 40 mM, more suitably of about 20 mM to 30 mM, totally suitably in a collective concentration of about 25 mM.

In one embodiment, the liquid pharmaceutical composition comprises a citrate buffer system comprising two or more buffer species, suitably selected from acid citrate, dihydrogen citrate, monohydrogen citrate and citrate (and / or derivatives thereof), suitably with counterions of sodium for any citrate species, at a collective concentration of about 1 to 50mM, suitably from about 10 to 40mM, more suitably from about 20mM to 30mM, most suitably at a collective concentration of about 25mM.

In one embodiment, when more than one buffer system is present (e.g., a citrate buffer system and a phosphate buffer system), the sum of the individual concentrations of each of the respective buffer systems (i.e., the collective concentration ) can suitably fall within any one of the above-mentioned buffer system concentrations.

Suitably, the liquid pharmaceutical composition comprises a phosphate buffer system, wherein the phosphate buffer species are present within the composition at a concentration (defined with reference to phosphoric acid equivalents, i.e., hypothetically assuming that all species buffer are in the form of phosphoric acid, MW = 98) from about 2.45 pg / ml to 24.5 mg / ml (i.e. 2.45 pg / ml to 24.5 mg / ml phosphoric acid equivalents ), suitably from about 0.245 mg / ml to about 12.25 mg / ml, suitably from about 1.225 mg / ml to about 3.675 mg / ml, totally suitably about 2.45 mg / ml.

Suitably, the weight ratio of phosphate buffer species (in terms of phosphoric acid equivalents) to etanercept within the liquid pharmaceutical composition is between 1: 200 and 1: 2, suitably between 1: 100 and 1: 4, suitably between 1:50 and 1:10, suitably around 1:20, 4.

Suitably, the liquid pharmaceutical composition comprises a citrate buffer system, wherein the citrate buffer species are present within the composition at a concentration (defined with reference to citric acid equivalents, i.e., hypothetically assuming that all species buffer are in the form of citric acid, MW = 192.12 g mol-1) of approximately 4.8 pg / ml to 48 mg / ml (i.e. 4.8 pg / ml to 48 mg / ml equivalents of citric acid), suitably from about 0.48 mg / ml to about 24 mg / ml, suitably from about 2.4 mg / ml to about 7.2 mg / ml, fully suitably around 4.8 mg / ml .

Suitably, the weight ratio of buffer species of citrate (in terms of citric acid equivalents) to etanercept within the liquid pharmaceutical composition is between 1: 100 and 1: 1, suitably between 1: 50 and 1: 2, suitably between 1:25 and 1: 5, suitably around 1:10.

Suitably, the liquid pharmaceutical composition comprises the buffer system (suitably a phosphate and / or citrate buffer system) in a molar ratio of buffer system (in terms of moles of buffer species) to etanercept of about 5: 1 to about 100: 1. In one embodiment, the buffer system is present in a molar ratio of buffer system to etanercept of from about 10: 1 to about 50: 1, most suitably about 25: 1. This includes when the or The "buffer agent (s)" (eg, sodium citrate) are formed by adding a strong base (eg, sodium hydroxide) to the conjugated acid of the buffering agent (eg, citric acid).

As illustrated in the Examples section, the liquid pharmaceutical compositions of the invention that include an acetate buffer system perform particularly well in stress tests, thus validating the stability and viability of the pharmaceutical product of the compositions of the invention. .

Tonicity regulator

The liquid pharmaceutical composition of the invention suitably comprises a "tonicity modifier" (or "tonicity regulator") or one or more tonicity regulators, suitably as defined herein.

The inclusion of a tonicity regulator adequately contributes to (or increases) the overall osmolality and osmolarity of the composition. Suitably, a tonicity regulator is present within the composition in an amount or concentration sufficient for the composition to be (substantially) isotonic with body fluids. However, the tonicity regulator (suitably in combination with the stabilizer) is also potentially important in stabilizing etanercept.

Any suitable tonicity regulator can be used. However, the tonicity regulator is selected suitably among the group including water soluble metal salts (e.g. sodium chloride, potassium chloride, magnesium chloride, calcium chloride), water soluble tonicity regulating sugars / sugar alcohols (although different from any component that serve as a stabilizer) and / or other water soluble polyols. Suitably, the tonicity regulator (s) are not buffering (ie, they give rise to little or no buffering effect). As such, any metal salt tonicity regulator is suitably not a buffering agent.

The liquid pharmaceutical composition may comprise one or more tonicity regulators, although preferably only a single "tonicity regulator" is present (notwithstanding any tonicity regulating effect imparted to the composition by components intended to fulfill another function as defined by the present specification, as a stabilizer).

Fully suitably the tonicity regulator is or comprises a metal salt (preferably a non-buffering water soluble metal salt). Suitably said metal salt is or comprises a metal halide, suitably an alkali or alkaline earth metal halide, suitably an alkali metal chloride.

In a particular embodiment, the tonicity regulator is or comprises sodium chloride. In a particular embodiment, the tonicity regulator is sodium chloride. Sodium chloride is a particularly advantageous tonicity regulator for use in conjunction with a sucrose or trehalose stabilizer, especially with a phosphate or citrate buffer system.

Suitably, the liquid pharmaceutical composition comprises the tonicity regulator (s) (more suitably sodium chloride) in a concentration of from about 10 to about 200 mM, more suitably from about 20 to about 150 mM, more suitably from about 25 to about 75 mM or from about 75 mM to about 125 mM. In one embodiment, the tonicity regulator (s) are present at a concentration between 40 and 60 mM, fully suitably around 50 mM. One embodiment has sodium chloride at a concentration of 50 mM. In one embodiment, the tonicity regulator (s) are present in a concentration between 90 and 110 mM, fully suitably around 100 mM. One embodiment features sodium chloride at a concentration of 100 mM.

Suitably, the liquid pharmaceutical composition comprises the tonicity regulator (s) (more suitably sodium chloride) in a concentration from about 0.58 mg / ml to about 11.6 mg / ml, more suitably from about 1.16 mg. / ml to about 8.76 mg / ml, more suitably from about 1.46 mg / ml to about 4.38 mg / ml or from about 4.38 mg / ml to about 7.3 mg / ml. In one embodiment, the tonicity regulator (s) are present in a concentration between 2.34 mg / ml and 3.50 mg / ml, fully suitably about 2.92 mg / ml. One embodiment features sodium chloride at a concentration of 2.92 mg / ml. In one embodiment, the tonicity regulator (s) are present in a concentration between 5.26 mg / ml and 6.42 mg / ml, more suitably around 5.84 mg / ml. One embodiment features sodium chloride at a concentration of 5.84 mg / ml.

Suitably, the liquid pharmaceutical composition comprises the tonicity regulator (s) (fully suitably sodium chloride) in a molar ratio of tonicity regulator to etanercept of from about 10: 1 to about 200: 1, more suitably from about 20: 1 to about 150: 1, fully suitably about 25: 1 to about 75: 1 or about 75: 1 to about 125: 1. In one embodiment, the tonicity regulator (s) are present in a molar ratio of tonicity regulator to etanercept of from about 40: 1 to about 60: 1, fully suitably about 50: 1. One embodiment exhibits sodium chloride in a molar ratio of sodium chloride to etanercept of about 50: 1. In one embodiment, the tonicity regulator (s) are present in a molar ratio of tonicity regulator to etanercept from about 90: 1 to about 110: 1, most suitably from about 100: 1. One embodiment displays sodium chloride in a molar ratio of sodium chloride to etanercept of about 100: 1.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that include a tonicity regulator as defined herein perform particularly well in stress tests, with compositions comprising chloride having particularly good performance. sodium in a range of amounts as stipulated.

Stabilizer

Suitably, the liquid pharmaceutical composition comprises a stabilizer, more suitably a sugar stabilizer. Suitably, said component facilitates the maintenance of the structural integrity of the biopharmaceutical drug, in particular during freezing and / or lyophilization and / or storage (especially when exposed to stress).

The liquid pharmaceutical composition may comprise one or more sugar stabilizers, although in preferred embodiments only a single sugar stabilizer is present.

Suitably the sugar stabilizer is a disaccharide.

The sugar stabilizer is sucrose and / or trehalose.

In a particular embodiment, the sugar stabilizer is sucrose.

Suitably, the liquid pharmaceutical composition comprises at most one sugar stabilizer, suitably at most one sugar and / or disaccharide polyol. Suitably, the liquid pharmaceutical composition comprises sucrose as the sole sugar stabilizer.

Suitably, the sucrose used to form the liquid pharmaceutical composition is anhydrous sucrose. Regardless of the hydration state used, any amount stipulated in relation to sucrose (unless otherwise indicated) corresponds to pure anhydrous sucrose. These amounts can be converted into a sucrose hydrate amount by applying an appropriate multiplier. Furthermore, in order to assess whether a given formulation falls within the scope of any of the definitions of amount of sucrose given herein, an amount of sucrose hydrate can be easily converted to a corresponding amount of anhydrous pure sucrose (with an equal number of moles) by applying said multiplier in reverse. This principle can be adopted for any sugar stabilizer component. Obviously, the concentrations, when given as molar concentration, will be the same regardless of the hydration state of the sugar stabilizer.

Suitably, the liquid pharmaceutical composition comprises the sugar stabilizer (s) (fully suitably sucrose) in a concentration by weight of from about 0.1 to about 10% by weight, more suitably from about 0.5 to about 5% by weight, more suitably from about 0.75 to about 1.25% by weight or from about 3.5 to about 4.5% by weight. In one embodiment, the sugar stabilizer (s) are present in a concentration by weight of about 1% by weight. In one embodiment, the sugar stabilizer (s) are present in a concentration by weight of about 4% by weight.

Suitably, the liquid pharmaceutical composition comprises the sugar stabilizer (s) (wholly suitably sucrose) in a concentration of from about 5 to about 200 mM, more suitably from about 10 to about 150 mM, more suitably from about 20 to about 40 mM or about 110 to about 130 mM. In one embodiment, the sugar stabilizer (s) are present in a concentration of about 30 mM. In one embodiment, the sugar stabilizer (s) are present in a concentration of 120 mM.

Suitably, the liquid pharmaceutical composition comprises the sugar stabilizer (s) (fully suitably sucrose) in a concentration from about 1 mg / ml to about 100 mg / ml, more suitably from about 5 mg / ml to about 50 mg / ml, more suitably from about 7.5 mg / ml to about 12.5 mg / ml or from about 35 mg / ml to about 45 mg / ml. In one embodiment, the sugar stabilizer (s) are present in a concentration of about 10 mg / ml. In a particular embodiment, the sugar stabilizer or stabilizers are present in a concentration of approximately 40 mg / ml.

Suitably, the liquid pharmaceutical composition comprises the sugar stabilizer (s) (fully suitably sucrose) in a molar ratio of sugar stabilizer (s) to etanercept of from about 10: 1 to about 200: 1, more suitably from about 20: 1 to about 150: 1, more suitably from about 25: 1 to about 125: 1. In one embodiment, the sugar stabilizer (s) are present in a molar ratio of sugar stabilizer (s) to etanercept from about 20: 1 to about 40: 1 or from about 110: 1 to about 130: 1. In one embodiment, the sugar stabilizer (s) are present in a molar ratio of sugar stabilizer (s) to etanercept. of about 30: 1. In one embodiment, the sugar stabilizer (s) They are present in a molar ratio of sugar stabilizer (s) to etanercept of about 120: 1.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that include a sugar stabilizer as defined herein perform particularly well in stress tests. Furthermore, liquid pharmaceutical compositions comprising sucrose as a sugar stabilizer work particularly well.

Polar ionizable amino acids ( and optional additional amino acids)

The liquid pharmaceutical composition suitably comprises a polar ionizable amino acid, suitably excluding arginine (and also suitably excluding any amino acid containing sulfur). The polar ionizable amino acid is suitably a naturally occurring amino acid. The polar ionizable amino acid is both a polar (or hydrophilic) amino acid and an ionizable (or charged) amino acid. These amino acid categorizations are well known in the art. The term "polar amino acid" refers to amino acids that carry a polar side chain, while the term "ionizable amino acid" refers to amino acids that carry a side chain that is or is capable of ionization (eg, protonation or deprotonation). Therefore, ionizable amino acids are acidic amino acids or basic amino acids.

For the purposes of the present disclosure, natural polar amino acids suitably include: arginine, asparagine, aspartic acid, glutamine, glutamic acid, histidine, lysine, serine, and threonine. Ionizable natural amino acids suitably include: aspartic acid, glutamic acid, histidine, cysteine, lysine, tyrosine, and arginine. Therefore, the polar ionizable natural amino acids include: aspartic acid and glutamic acid, which can be considered "acidic amino acids"; and histidine, lysine, and arginine, which can be considered "basic amino acids." Thus, suitably, the polar ionizable amino acid is selected from the group consisting of aspartic acid, glutamic acid, histidine, and lysine.

In a particular embodiment, the polar ionizable amino acid is selected from the group consisting of aspartic acid, histidine, and lysine.

When the polar ionizable amino acid is a basic amino acid, especially one of the more strongly basic amino acids (that is, when the pK ^a of the acid / protonated side chain conjugate is greater than or equal to 10.0, thus excluding histidine ), the liquid pharmaceutical composition may suitably comprise an additional non-polar amino acid, suitably excluding arginine (and also suitably excluding any sulfur-containing amino acids). The nonpolar amino acid is suitably a naturally occurring amino acid. The term "non-polar amino acid" refers to amino acids that carry a non-polar (or hydrophobic) side chain. For the purposes of the present disclosure, natural non-polar amino acids suitably include: alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine, and valine.

Suitably, the nonpolar amino acid is one which a-amino group has a pK value relatively high (ie, with respect to the pK ^a of its conjugate acid / protonated form), suitably a pK ^a value that is similar to the pK of the side chain of its accompanying basic amino acid. Suitably the pK ^a of the acid conjugate / protonated form of the a-amino group of the non-polar amino acid is greater than or equal to 10.0). In a particular embodiment, the non-polar amino acid is proline.

Suitably, when the liquid pharmaceutical composition comprises both a polar ionizable amino acid and a non-polar amino acid (eg, lysine and proline), any amount, concentration, or ratio stipulated herein in relation to the polar ionizable amino acid can only represent Total amounts, concentrations, or proportions of the polar ionizable amino acid and the non-polar amino acid in combination. As such, a non-polar amino acid can be included to replace a portion of the polar ionizable amino acid. The combination of lysine and proline is especially feasible in this regard, particularly in certain preferred proportions.

When the liquid pharmaceutical composition comprises both a polar ionizable amino acid and a non-polar amino acid (eg, lysine and proline), suitably the molar ratio of polar ionizable amino acid to non-polar amino acid is between 10: 1 and 1: 10, plus suitably between 5: 1 and 1: 5, more suitably between 2: 1 and 1: 2, totally suitably about 1: 1 (suitably / - 10%). As such, in a particular embodiment, the liquid pharmaceutical composition comprises lysine and proline in one of the above-mentioned molar ratios, more suitably 1: 1, and suitably the total amounts, concentrations or ratios (for example, relative to another non-component amino acid) of the two amino acids combined are defined by the amounts, concentrations, or ratios set forth herein relative to the polar ionizable amino acid alone.

Suitably, therefore, the liquid pharmaceutical composition suitably comprises a polar ionizable amino acid selected from the group consisting of:

- Aspartic acid;

- histidine; Y

- lysine (optionally and fully suitably in combination with a non-polar amino acid, fully suitably proline).

Suitably, provided that the liquid pharmaceutical composition comprises a polar ionizable basic amino acid with a side chain whose acid / protonated conjugate has a pK ^at greater than or equal to 10, the liquid pharmaceutical composition further comprises a non-polar amino acid (suitably excluding any amino acid containing sulfur). Suitably, no more amino acids are present.

Suitably, provided that the liquid pharmaceutical composition comprises a polar ionizable amino acid with a side chain whose acid / protonated conjugate has a pK ^of less than 10, the liquid pharmaceutical composition is (substantially or totally) free of any non-polar amino acids. , and suitably is (substantially or totally) free of any other amino acid.

Suitably, the liquid pharmaceutical composition suitably comprises:

- Aspartic acid;

- histidine; or

- a combination of lysine and proline;

suitably in an amount, concentration, or ratio (eg, relative to a component other than an amino acid) defined by any of the amounts, concentrations, or proportions set forth herein relative to the polar ionizable amino acid alone.

Suitably, the liquid pharmaceutical composition comprises a polar ionizable amino acid (optionally in combination with a non-polar amino acid as defined above) in a concentration of from about 1 to 50 mM, more suitably from about 10 to 40 mM, so fully adequate from about 20 mM to 30 mM. In a particular embodiment, the liquid pharmaceutical composition comprises a polar ionizable amino acid (optionally in combination with a non-polar amino acid as defined above) in a concentration of approximately 25 mM.

Suitably, the liquid pharmaceutical composition comprises the polar ionizable amino acid (optionally in combination with a non-polar amino acid as defined above) in a polar ionizable amino acid molar ratio (optionally in combination with a non-polar amino acid as defined above) relative to etanercept from about 5: 1 to about 100: 1. In one embodiment, the polar ionizable amino acid (optionally in combination with a non-polar amino acid as defined above) is present in a molar amino acid ratio ionizable polar (optionally in combination with a non-polar amino acid as defined above) to etanercept from about 10: 1 to about 50: 1, more suitably about 25: 1.

The amino acid (s) described in this section may suitably be provided in the zwitterionic form or in the salt form (eg, HCl salt). Suitably, unless otherwise indicated, any amount or concentration (either absolute or relative) given herein in relation to amino acids suitably refers to the amounts or concentrations of the respective free hybrid amino acids, even though said amino acids they may be provided in a different form or may even exist within the liquid pharmaceutical composition in a different form with the prevailing pH.

Thinner

The liquid pharmaceutical compositions of the invention may include one or more pharmaceutically acceptable diluents. However, fully suitably the liquid pharmaceutical composition is an aqueous pharmaceutical composition. Suitably the diluent consists of water and suitably water alone. The water is suitably Water for Injection (WFI).

Suitably, the diluent can constitute the rest of the ingredients in any liquid pharmaceutical composition, for example, so that the percentages by weight add up to 100%. Suitably, any concentration given herein in relation to any component of the liquid pharmaceutical composition represents concentrations of that component in (and suitably dissolved in) the diluent in admixture with any other component.

The liquid pharmaceutical composition of the invention is suitably a solution and is suitably (substantially or totally) free of particles or precipitates.

Components not present or low level

Low / no arginine content

Suitably the liquid pharmaceutical composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of arginine or comprises arginine in a molar ratio of arginine to buffer system of at most 1: 150 (that is, less than or equal to one mole of arginine for each 150 moles of buffer system), more suitably at most 1: 1,500, fully suitably at most 1: 15,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of arginine or comprises arginine in a weight ratio of arginine to etanercept of at most 1: 3,000 (i.e. less than or equal to one part by weight of arginine per 3,000 parts per etanercept), more suitably at most 1: 30,000, fully adequately at most 1: 300,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of arginine or comprises arginine in a molar ratio of arginine to etanercept of at most 1: 3.75 (i.e. less than or equal to one mole of amino acid (s) for every 3.75 moles of etanercept), more suitably at most 1: 37.5, fully suitably at most 1: 375.

As explained herein, such references to "amino acids" (in this case arginine) in the context of their presence or otherwise within liquid pharmaceutical compositions refer to the corresponding free amino acid (s) ( s) and not to amino acid residue (s) covalently incorporated as part of a larger compound, such as a peptide or protein.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that exclude (substantially or totally) arginine perform particularly well in stress tests and are much more stable with respect to protein aggregation, fragmentation and unfolding. than expected in view of the prior art.

Amino acid content with low / no sulfur content

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of sulfur-containing amino acid (s) (e.g. cysteine or methionine, especially methionine) or comprises sulfur-containing amino acid (s) (e.g. , cysteine or methionine, especially methionine) in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of sulfur-containing amino acid (s) (e.g. cysteine or methionine, especially methionine) or comprises sulfur-containing amino acid (s) (e.g. , cysteine or methionine, especially methionine) in a molar ratio of amino acid (s) containing sulfur to buffer system of at most 1: 150 (that is, less than or equal to one mole of amino acid (s) containing (s) sulfur per 150 moles of buffer system), more suitably at most 1: 1,500, fully suitably at most 1: 15,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of sulfur-containing amino acid (s) (e.g. cysteine or methionine, especially methionine) or comprises sulfur-containing amino acid (s) (e.g. , cysteine or methionine, especially methionine) in a weight ratio of amino acid (s) containing sulfur to etanercept of at most 1: 3,000 (i.e. less than or equal to one part by weight of amino acid ( s) containing sulfur per 3,000 parts by weight of etanercept), more suitably at most 1: 30,000, fully suitably at most 1: 300,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of sulfur-containing amino acid (s) or comprises sulfur-containing amino acid (s) in a molar ratio of sulfur-containing amino acid (s). sulfur to etanercept of at most 1: 3.75 (i.e. less than or equal to one mole of amino acid (s) for every 3.75 moles of etanercept), more suitably at most 1: 37.5, totally adequately at most 1: 375.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of cysteine and / or methionine (totally preferably both), or comprises cysteine and / or methionine (totally preferably both) in an amount, concentration, molar ratio or weight ratio of at most that stipulated in any of the preceding paragraphs of this subsection in relation to "amino acid (s) containing sulfur" more generally.

As explained herein, such references to "amino acids" (in this case, sulfur-containing amino acid (s)) in the context of their presence or otherwise within liquid pharmaceutical compositions refer to or to the corresponding free amino acid (s) and not to amino acid residue (s) covalently incorporated as part of a larger compound, such as a peptide or protein.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that exclude (substantially or totally) sulfur-containing amino acid (s) (eg, cysteine or methionine, especially methionine) perform particularly well in stress tests.

Low / no amino acid content not stipulated

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of non-stipulated amino acid (s) (i.e. amino acids other than those stipulated as present) or comprises non-stipulated amino acid (s) in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM.

As explained herein, such references to "amino acids" (in this case non-stipulated amino acids) in the context of their presence or otherwise within liquid pharmaceutical compositions refer to the free amino acid (s) ( s) corresponding to and not to amino acid residue (s) covalently incorporated as part of a larger compound, such as a peptide or protein.

Low / no surfactant content

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of surfactants (either cationic, anionic, amphoteric or non-ionic) or comprises one or more of said surfactants in a concentration (collective) of at most 1 mM, more suitably at most 0.1 mM, more suitably at most 0.01 mM, more suitably at most 0.001 mM, fully suitably at most 0.0001 mM.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of surfactants (whether cationic, anionic, amphoteric or nonionic) or comprises one or more of said surfactants in a (collective) molar ratio of surfactant (s) to buffer system of at most 1: 10, more suitably at most 1: 100, fully suitably at most 1: 1,000, most suitably at most 1: 10,000, suitably at most 1: 100,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of surfactants (whether cationic, anionic, amphoteric or nonionic) or comprises one or more of said surfactants in a (collective) weight ratio of surfactant (s) to to etanercept of at most 1: 50 (i.e. less than or equal to one part by weight of surfactant (s) for every 50 parts by weight of etanercept), more suitably at most 1: 500, more suitably at most 1: 5,000, more suitably 1: 50,000 at most, suitably 1: 500,000 at most.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of surfactants (whether cationic, anionic, amphoteric or nonionic) or comprises one or more of said surfactants in a (collective) molar ratio of surfactant (s) to etanercept of at most 3: 1, more suitably at most 0.3: 1, more suitably 0.003: 1, more suitably 0.0003: 1, suitably 0.00003: 1.

Suitably, the surfactant (s) referred to in this section (and therefore considered non-existent or present only in very low amounts) may be cationic, anionic, amphoteric or non-ionic surfactants. Suitably, the surfactant (s) mentioned in this section (and which are considered non-existent or present only in very low amounts) are non-ionic surfactants (eg, polysorbates or ranges). Therefore, the liquid pharmaceutical composition is suitably (substantially or totally) free of nonionic surfactants (especially polysorbate 20 and polysorbate 80), or comprises one or more of said nonionic surfactants in an amount, concentration, molar ratio or ratio in weight of at most that stipulated in any of the previous paragraphs of this subsection in relation to "surfactant (s)" more generally.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that exclude (substantially or entirely) surfactants or certain surfactants, as defined above, perform particularly well in stress testing. This may be considered surprising in view of the prior art, which often employs a surfactant when arginine is absent.

Low / no nitrogen chelating agents

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (e.g. EDTA or salts thereof) or comprises chelating agent (s) containing (n) Nitrogen (eg EDTA or salts thereof) in a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (e.g. EDTA or salts thereof) or comprises chelating agent (s) containing Nitrogen (for example EDTA or salts thereof) in a molar ratio of chelating agent (s) containing nitrogen (for example EDTA or salts thereof) to a buffer system at most 1: 150 (that is, less than or equal to one mole of chelating agent (s) containing (s) nitrogen for every 150 moles of buffer system), more suitably at most 1: 1,500, so fully adequate at most 1: 15,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (e.g. EDTA or salts thereof) or comprises chelating agent (s) containing (n) nitrogen (eg EDTA or salts thereof) in a weight ratio of chelating agent (s) containing nitrogen (eg EDTA or salts thereof) to etanercept as at most 1: 3,000 (i.e. less than or equal to one part by weight of chelating agent (s) containing (s) nitrogen per 3,000 parts by weight of etanercept), more suitably at most 1: 30,000, of fully adequate form at most 1: 300,000.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (e.g. EDTA or salts thereof) or comprises chelating agent (s) containing Nitrogen (for example EDTA or salts thereof) in a molar ratio of chelating agent (s) containing nitrogen (for example EDTA or salts thereof) to etanercept at the most 1: 3.75 (i.e. less than or equal to one mole of chelating agent (s) containing (s) nitrogen for every 3.75 moles of etanercept), most suitably at most 1: 37.5 , fully adequately at most 1: 375.

Suitably, the liquid pharmaceutical composition is (substantially or totally) free of EDTA or a salt thereof, or comprises EDTA or a salt thereof in an amount, concentration, molar ratio or weight ratio of at most that set forth in any of the preceding paragraphs of this subsection regarding "chelating agent (s) containing (s) nitrogen" more generally.

Such references to "nitrogen-containing chelating agent (s)" in the context of their presence or otherwise within liquid pharmaceutical compositions refer to the free chelating agent (s) ( s) and not to chelating residues covalently incorporated as part of a peptide or protein.

As illustrated in the Examples section, liquid pharmaceutical compositions of the invention that exclude (substantially or totally) the nitrogen-containing chelating agent (s) (eg, EDTA) perform particularly well. good on stress tests.

Particular achievements

In a particular embodiment, the liquid pharmaceutical composition comprises a citrate buffer system and either histidine or aspartic acid as polar ionizable amino acid. In a particular embodiment, the liquid pharmaceutical composition comprises a buffer system of citrate and histidine as a polar ionizable amino acid.

In a particular embodiment, the liquid pharmaceutical composition comprises a phosphate buffer system and: it is (substantially or completely) free of any amino acid; comprises aspartic acid (and suitably no other amino acids); or comprises a combination of lysine and proline (and suitably no other amino acids). The molar concentrations of the stabilizer (for example, sucrose) and the tonicity regulator (for example, sodium chloride) can be interrelated by Equation (1):

[stabilizer] ^moi * [tonicity regulator] ^moi = A ^mol * 2n Equation (1)

where [stabilizer] ^moi is the molar concentration of the stabilizer; [tonicity regulator] ^moi is the molar concentration of the tonicity regulator; A ^moi is a reference molar concentration constant (the product of molar concentrations when n = 0); and n is any number (positive or negative).

The concentrations by weight of the stabilizer (for example, sucrose) and the tonicity regulator (for example, sodium chloride) can be interrelated by Equation (2):

[stabilizer] ^weight * [tonicity regulator] ^weight = A ^weight * 2n Equation (2)

where [stabilizer] ^weight is the weight concentration of the stabilizer; [tonicity regulator] ^weight is the concentration by weight of the tonicity regulator; A ^weight is a reference weight concentration constant (the product of the weight concentrations when n = 0); and n is any number (positive or negative).

In relation to Equation (1), suitably A ^moi is between 1,000 and 5,000 mM2 (units = square millimoles, mM2), suitably between 2,000 and 4,000 mM2, more suitably between 2,900 and 3,100 mM2, fully suitably around 3,000 mM2 .

In relation to equation (1), suitably n is a number between -5 and 5, suitably a number between -2 and 2, more suitably a number between -1 and 1, totally suitably 0 or 1.

In relation to Equation (2), suitably A ^weight is between 20 and 100 (mg / ml) 2 (units = square milligrams per milliliter, (mg / ml) 2), suitably between 40 and 80 (mg / ml) 2 , more suitably between 50 and 70 (mg / ml) 2, totally suitable around 58 (mg / ml) 2.

In relation to equation (2), suitably n is a number between -5 and 5, suitably a number between -2 and 2, more suitably a number between -1 and 1, totally suitably 0 or 1.

As an example, the molar weights and the concentrations of sucrose (stabilizer) and sodium chloride (tonicity regulator) can be interrelated as follows:

where A ^moi is 3,000 mM2 and A ^wt is 58 (mg / ml) 2.

Varying the molar or weight concentrations of the sugar and salt in accordance with Equations (1) and / or (2) generally produces liquid pharmaceutical compositions exhibiting viable performance and storage characteristics.

In preferred embodiments, the minimum stabilizer weight concentration is 0.5% by weight. In realizations preferred, the minimum molar concentration of stabilizer is 15 mM. In preferred embodiments, the minimum stabilizer weight concentration is 5 mg / ml.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or wholly) free of amino acid (s) or comprises (preferably as substantially or completely the only amino acid (s)) a polar ionizable natural amino acid and optionally an additional non-polar amino acid.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or wholly) free of amino acid (s) or comprises (preferably as substantially or completely the only amino acid (s)) a polar ionizable natural amino acid and optionally an additional non-polar amino acid;

wherein the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM; wherein the composition is (substantially or totally) free of sulfur-containing amino acid (s) (especially methionine) or comprises sulfur-containing amino acid (s) in a concentration of not more than 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM;

wherein the composition is (substantially or totally) free of surfactants (especially non-ionic surfactants such as polysorbates and ranges) or comprises one or more of said surfactants in a (collective) concentration of at most 1 mM, more suitably at most 0 , 1 mM, most suitably at most 0.01 mM, most suitably at most 0.001 mM, most suitably at most 0.0001 mM; Y

wherein the composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (especially EDTA or salts thereof) or comprises nitrogen-containing chelating agent (s) at a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

where the molar concentrations of the stabilizer (for example, sucrose) and the tonicity regulator (for example example, sodium chloride) are interrelated by Equation (1):

[stabilizer] ^moi * [tonicity regulator \ ^moi = A ^moi * 2n Equation (1)

where [stabilizer] ^moi is the molar concentration of the stabilizer; [tonicity regulator] ^moi is the molar concentration of the tonicity regulator; A ^moi is a reference molar concentration constant between 2,000 and 4,000 mM2; and n is a number between -2 and 2;

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only or the only amino acids) a polar ionizable natural amino acid in a concentration between 10 and 30 mM and optionally a non-polar amino acid additional in a concentration between 10 and 30 mM.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5;

where the molar concentrations of the stabilizer (for example, sucrose) and the tonicity regulator (for example, sodium chloride) are interrelated by Equation (1):

[stabilizer] ^moi * [tonicity regulator] ^moi = A ^moi * 2n Equation (1)

where [stabilizer] ^moi is the molar concentration of the stabilizer; [tonicity regulator] ^moi is the molar concentration of the tonicity regulator; A ^moi is a reference molar concentration constant between 2,000 and 4,000 mM2; and n is a number between -2 and 2;

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only or the only amino acids) a polar ionizable natural amino acid in a concentration between 10 and 30 mM and optionally a non-polar amino acid additional in a concentration between 10 and 30 mM; wherein the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM; wherein the composition is (substantially or totally) free of sulfur-containing amino acid (s) (especially methionine) or comprises sulfur-containing amino acid (s) in a concentration of not more than 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM;

wherein the composition is (substantially or totally) free of surfactants (especially non-ionic surfactants such as polysorbates and ranges) or comprises one or more of said surfactants in a (collective) concentration of at most 1 mM, more suitably at most 0 , 1 mM, most suitably at most 0.01 mM, most suitably at most 0.001 mM, most suitably at most 0.0001 mM; Y

wherein the composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (especially EDTA or salts thereof) or comprises nitrogen-containing chelating agent (s) at a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- aspartic acid (preferably with a phosphate buffer);

- histidine (preferably with a citrate buffer); or

- a combination of lysine and proline.

In one embodiment, the liquid pharmaceutical composition comprises:

- etanercept;

- a buffer system (for example, phosphate buffer or citrate buffer);

- a tonicity regulator (for example sodium chloride);

- a sugar stabilizer (eg sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- aspartic acid (preferably with a phosphate buffer);

- histidine (preferably with a citrate buffer); or

- a combination of lysine and proline; Y

wherein the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM; wherein the composition is (substantially or totally) free of sulfur-containing amino acid (s) (especially methionine) or comprises sulfur-containing amino acid (s) in a concentration of not more than 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM;

wherein the composition is (substantially or totally) free of surfactants (especially non-ionic surfactants such as polysorbates and ranges) or comprises one or more of said surfactants in a (collective) concentration of at most 1 mM, more suitably at most 0 , 1 mM, most suitably at most 0.01 mM, most suitably at most 0.001 mM, most suitably at most 0.0001 mM; Y

wherein the composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (especially EDTA or salts thereof) or comprises nitrogen-containing chelating agent (s) at a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

In one embodiment, the liquid pharmaceutical composition comprises:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer or citrate buffer);

40-120 mM tonicity regulator (eg sodium chloride) (suitably 40-60 mM or 90-110 mM); - 0.5-5.0% by weight of sugar stabilizer (for example sucrose and / or trehalose) (suitably 0.5 to 1.5% by weight or 3.5 to 4 , 5% by weight); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline.

In one embodiment, the liquid pharmaceutical composition comprises:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer or citrate buffer);

40-120 mM tonicity regulator (eg sodium chloride) (suitably 40-60 mM or 90-110 mM); - 0.5-5.0% by weight of sugar stabilizer (for example sucrose and / or trehalose) (suitably 0.5 to 1.5% by weight or 3.5 to 4 , 5% by weight); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline; Y

wherein the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM; wherein the composition is (substantially or totally) free of sulfur-containing amino acid (s) (especially methionine) or comprises sulfur-containing amino acid (s) in a concentration of not more than 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM;

wherein the composition is (substantially or totally) free of surfactants (especially non-ionic surfactants such as polysorbates and ranges) or comprises one or more of said surfactants in a (collective) concentration of at most 1 mM, more suitably at most 0 , 1 mM, most suitably at most 0.01 mM, most suitably at most 0.001 mM, most suitably at most 0.0001 mM; Y

wherein the composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (especially EDTA or salts thereof) or comprises nitrogen-containing chelating agent (s) at a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer or citrate buffer);

- 40-120 mM tonicity regulator (sodium chloride);

- 0.5-5.0% by weight of sugar stabilizer (for example, sucrose and / or trehalose); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer);

- tonicity regulator 90-110 mM (for example, sodium chloride);

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sugar stabilizer (for example, sucrose and / or trehalose); and - water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer);

- tonicity regulator 90-110 mM (for example, sodium chloride);

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sugar stabilizer (for example, sucrose and / or trehalose); - 20-30 mM aspartic acid; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg citrate buffer);

- tonicity regulator 90-110 mM (for example, sodium chloride);

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sugar stabilizer (for example, sucrose and / or trehalose); - 20-30 mM histidine; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system (eg phosphate buffer);

- 45-55 mM tonicity regulator (for example, sodium chloride);

- 3.5-4.5% by weight (or 120 mM / -30 mM) of sugar stabilizer (for example, sucrose and / or trehalose); - 10-15 mM lysine;

- 10-15 mM proline; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM buffer system (eg phosphate buffer);

- 100 mM tonicity regulator (for example, sodium chloride);

- 1% by weight (or 30 mM / -1 mM) of sugar stabilizer (for example, sucrose and / or trehalose); and - water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM buffer system (eg phosphate buffer);

- 100 mM tonicity regulator (for example, sodium chloride);

- 1% by weight (or 30 mM / -1 mM) of sugar stabilizer (for example, sucrose and / or trehalose);

- 25 mM aspartic acid; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM buffer system (eg citrate buffer);

- 100 mM tonicity regulator (for example, sodium chloride);

- 1% by weight (or 30 mM / -1 mM) of sugar stabilizer (for example, sucrose and / or trehalose);

- 25 mM histidine; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM buffer system (eg phosphate buffer);

- 50 mM tonicity regulator (for example, sodium chloride);

- 4% by weight (or 120 mM / -4 mM) of sugar stabilizer (for example sucrose and / or trehalose);

- 12.5 mM lysine;

- 12.5 mM proline; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition comprises:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system selected from a phosphate buffer system or a citrate buffer system; - 40-120 mM sodium chloride (suitably 40-60 mM or 90-110 mM);

- 0.5-5.0% by weight of sucrose (suitably 0.5 to 1.5% by weight or 3.5 to 4.5% by weight); and - water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline.

In one embodiment, the liquid pharmaceutical composition comprises:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system selected from a phosphate buffer system or a citrate buffer system; - 40-120 mM sodium chloride (suitably 40-60 mM or 90-110 mM);

- 0.5-5.0% by weight of sucrose (suitably 0.5 to 1.5% by weight or 3.5 to 4.5% by weight); Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline; Y

wherein the composition is (substantially or totally) free of arginine or comprises arginine in a concentration of at most 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM; wherein the composition is (substantially or totally) free of sulfur-containing amino acid (s) (especially methionine) or comprises sulfur-containing amino acid (s) in a concentration of not more than 0.1 mM, more suitably at most 0.01 mM, fully suitably at most 0.001 mM;

wherein the composition is (substantially or totally) free of surfactants (especially non-ionic surfactants such as polysorbates and ranges) or comprises one or more of said surfactants in a (collective) concentration of at most 1 mM, more suitably at most 0 , 1 mM, most suitably at most 0.01 mM, most suitably at most 0.001 mM, most suitably at most 0.0001 mM; Y

wherein the composition is (substantially or totally) free of nitrogen-containing chelating agent (s) (especially EDTA or salts thereof) or comprises nitrogen-containing chelating agent (s) at a concentration of at most 0.1mM, more suitably at most 0.01mM, fully suitably at most 0.001mM.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM buffer system selected from a phosphate buffer system or a citrate buffer system; - 40-120 mM sodium chloride;

- 0.5-5.0% by weight of sucrose; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5; Y

wherein the composition is (substantially or totally) free of amino acid (s) or comprises (preferably as substantially or totally the only amino acid (s)):

- 20-30 mM aspartic acid (preferably with a phosphate buffer);

- 20-30 mM histidine (preferably with a citrate buffer); or

- a combination of 10-15 mM lysine and 10-15 mM proline.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM phosphate buffer system (suitably sodium phosphate (s) as buffering agent);

- 90-110 mM sodium chloride;

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sucrose; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM phosphate buffer system (suitably sodium phosphate (s) as buffering agent); - 90-110 mM sodium chloride;

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sucrose;

- 20-30 mM aspartic acid; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM citrate buffer system (suitably sodium citrate (s) as a buffering agent); - 90-110 mM sodium chloride;

- 0.9-1.1% by weight (or 30 mM / -5 mM) of sucrose;

- 20-30 mM histidine; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 45-55 mg / ml (or alternatively 20-30 mg / ml) of etanercept;

- 20-30 mM phosphate buffer system (suitably sodium phosphate (s) as a buffering agent); - 45-55 mM sodium chloride;

- 3.5-4.5% by weight (or 120 mM / -30 mM) of sucrose;

- 10-15 mM lysine;

- 10-15 mM proline; Y

- water (for injection);

wherein the composition has a pH between 6.1 and 6.5.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM phosphate buffer system (suitably sodium phosphate (s) as a buffering agent); - 100 mM sodium chloride;

- 1% by weight (or 30 mM / -1 mM) of sucrose; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM phosphate buffer system (suitably sodium phosphate (s) as a buffering agent); - 100 mM sodium chloride;

- 1% by weight (or 30 mM / -1 mM) of sucrose;

- 25 mM aspartic acid; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM citrate buffer system (suitably sodium citrate (s) as a buffering agent);

- 100 mM sodium chloride;

- 1% by weight (or 30 mM / -1 mM) of sucrose;

- 25 mM histidine; Y

- water (for injection);

wherein the composition has a pH of 6.3.

In one embodiment, the liquid pharmaceutical composition consists essentially of:

- 50 mg / ml (or alternatively 25 mg / ml) of etanercept;

- 25 mM phosphate buffer system (suitably sodium phosphate (s) as a buffering agent);

- 50 mM sodium chloride;

- 4% by weight (or 120 mM / -4 mM) of sucrose;

- 12.5 mM lysine;

- 12.5 mM proline; Y

- water (for injection);

wherein the composition has a pH of 6.3.

Suitably, the liquid pharmaceutical composition may be as set forth in any of the preceding embodiments, except that, rather than being defined by the amounts and / or concentrations indicated above of particular components (regardless of whether they are present or in minute amounts ), are instead or additionally defined with reference to the corresponding molar or weight ratios of the component to the buffer system; or the corresponding molar or weight ratios of the component to etanercept. The person skilled in the art will readily deduce for each component, from the relevant section of this specification relating to that specific component, which molar and weight ratios correspond to which concentrations. It will be appreciated that the buffering agent or buffer system of any of the embodiments described herein can be incorporated directly into the compositions or can be produced in situ, for example, through an acid-base reaction, suitably reacting a conjugated acid (eg phosphoric or citric acid) with a base (eg sodium hydroxide); or a conjugated base / base (eg sodium phosphate species or sodium citrate species) with an acid (eg hydrochloric acid). Regardless of the method used to provide or produce the buffer agent or buffer system, suitably the resulting composition ultimately comprises an appropriate balance of buffering agent and any acid / base conjugate to provide the desired pH. The person skilled in the art will be able to easily calculate or determine experimentally, without undue effort, the appropriate balance of buffer agent and acid / base conjugate, and / or the amount of base that must be added to a conjugate acid to produce the appropriate amount of agent. buffer and provide the desired pH.

Method of preparing a liquid pharmaceutical composition

The present invention provides a method of preparing a liquid pharmaceutical composition (suitably as defined herein) which suitably includes mixing together the respective ingredients stipulated with respect to the liquid pharmaceutical composition, optionally in any amount, concentration or stipulated form; and optionally adjusting one or more parameters given herein in relation to a liquid pharmaceutical composition (eg, pH).

The method suitably comprises mixing together, in any particular order deemed appropriate, any relevant components required to form a liquid pharmaceutical composition as defined herein. The person skilled in the art can refer to Examples or techniques well known in the field for forming liquid pharmaceutical compositions (especially those for injection by syringe). Different embodiments will suitably require different combinations of components to be mixed, potentially in different amounts. The person skilled in the art can easily deduce said combinations and amounts with reference to the previous description related to the liquid pharmaceutical composition.

Suitably, the method includes mixing the relevant components together in a suitable manner, in a diluent (eg, water), suitably such that all components dissolve (substantially or totally) in the diluent.

The method may include first preparing a premix (or pre-solution) of some or all of the components (optionally with part of the diluent or with all of the diluent) excluding etanercept, and then the etanercept itself can be mixed (optionally with part of the diluent). diluent or previously dissolved therein) with the premix (or premix) to produce the liquid pharmaceutical composition, or a composition to which the final components are then added to provide the final liquid pharmaceutical composition. More suitably, the premix contains all of the components except the etanercept and optionally also some diluent (which can be used to pre-dissolve the etanercept), suitably to add the etanercept to a mixture that offers optimal stabilization of the etanercept. Suitably, the aforementioned premix is prepared at the desired pH for the final liquid pharmaceutical formulation.

Suitably, the method involves forming a buffer system, suitably a buffer system comprising a buffering agent as defined herein. The buffer system is suitably formed in a premix prior to the addition of etanercept, although the buffer system can optionally be formed with etanercept present. The buffer system can be formed simply by mixing the buffer agent (supplied ready to use) with its acid / base conjugate (suitably in appropriate relative amounts to provide the desired pH; this can be determined by one of ordinary skill in the art, whether theoretical or experimentally). In the case of a phosphate (or citrate) buffer system, this may mean mixing sodium phosphate (or sodium citrate) and phosphoric acid (or citric acid) species together. Alternatively, the buffer system can be formed by adding a strong acid (eg HCl) to the buffer agent (eg sodium phosphate species) to form the acid / base conjugate in situ (eg phosphoric acid or hydrogen phosphates. ) (again suitably in appropriate relative amounts to provide the desired pH). Alternatively, the buffer system can be formed by adding a strong base (e.g. sodium hydroxide) to the acid / base conjugate (e.g. phosphoric acid or hydrogen phosphate (s)) of the buffering agent (e.g. phosphate (s) ) sodium) to form the buffering agent in situ (again suitably in appropriate relative amounts to provide the desired pH). The pH of the final liquid pharmaceutical composition premix can be judiciously adjusted by adding the required amount of strong base or strong acid, or even an amount of buffer or acid / base conjugate.

In certain embodiments, the buffering agent and / or the buffer system are pre-formed as a separate mixture and the buffer system is transferred to a precursor of the liquid pharmaceutical composition (comprising some or all of the components except the buffering agent and / or the buffer system, suitably comprising etanercept and potentially only etanercept) via buffer exchange (eg, by diafiltration to the relevant concentrations or osmolality). Additional excipients can then be added if necessary to produce the final liquid pharmaceutical composition. The pH can be adjusted after or before all the components are present.

Any, some or all of the components can be dissolved or premixed with a diluent before mixing with other components.

The final liquid pharmaceutical composition can be filtered, suitably to remove particulate material. Suitably, the filtration is carried out through filters with a size equal to or less than 1 pm, suitably 0.22 pm. Suitably, filtration is done through PES filters or PVDF filters, suitably 0.22 pm PES filters.

The present invention also provides a liquid pharmaceutical composition obtainable, obtainable through, or directly obtainable by the method of manufacture described herein.

Drug delivery device

The present invention also provides a drug delivery device comprising a liquid pharmaceutical composition as defined herein. Suitably, the drug delivery device comprises a chamber within which the pharmaceutical composition is located. Suitably the drug delivery device is sterile.

The drug delivery device will consist of a vial, ampoule, syringe, injection pen (eg, essentially incorporating a syringe), or intravenous bag. More suitably, the drug delivery device consists of a syringe, suitably an injection pen. Suitably, the syringe is a glass syringe. Suitably the syringe comprises a needle, suitably a 29G 1V needle.

The present invention provides a method of manufacturing a drug delivery device, suitably as defined herein, the method comprising incorporating a liquid pharmaceutical composition as defined herein into a drug delivery device. . Such manufacture typically involves loading the liquid pharmaceutical composition as defined herein into a syringe, suitably through a needle attached thereto. Later, the needle can be removed, replaced or maintained.

According to an eleventh aspect of the present invention there is provided a drug delivery device obtainable, obtainable through or directly obtained by the manufacturing method described herein.

Container

The present invention also provides a container comprising a liquid pharmaceutical composition as defined herein. Suitably, the package comprises a drug delivery device as defined herein, suitably a plurality of drug delivery devices. The package can comprise any container suitable for containing one or more drug delivery devices.

The present invention provides a method of manufacturing a container, the method comprising incorporating a liquid pharmaceutical composition as defined herein into a container. Suitably, this is accomplished by incorporating said liquid pharmaceutical composition into one or more drug delivery device (s), and then incorporating the pre-filled drug delivery device (s) into a container present within the package.

The container may suitably comprise instructions for using the liquid pharmaceutical composition, suitably for one or more medical indications set forth herein.

The present invention provides a container obtainable, obtainable through, or directly obtainable by a manufacturing method defined herein.

Parts kit

The present invention also provides a kit of parts comprising a drug delivery device (without the liquid pharmaceutical composition incorporated therein), a liquid pharmaceutical composition as defined herein (optionally contained in a separate container or container ) and optionally a set of instructions with directions regarding the administration (eg, subcutaneous) of the liquid pharmaceutical composition. The user can fill the drug delivery device with the liquid pharmaceutical composition (which can be provided in a vial or ampoule or the like) prior to administration.

Uses of the liquid pharmaceutical composition and methods of treatment

The present invention also provides:

- a method of treating a medical disease or disorder, suitably an autoimmune disease related to tumor necrosis factor alpha (TNF-a), suitably a disease selected from rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, plaque psoriasis and / or ankylosing spondylitis;

- a liquid pharmaceutical composition for use in therapy, suitably in the treatment of an autoimmune disease related to tumor necrosis factor alpha (TNF-a), suitably a disease selected from rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, psoriasis in plaques and / or ankylosing spondylitis;

- a use of liquid pharmaceutical compositions in the manufacture of a medicament for the treatment of a disease or disorder, suitably an autoimmune disease related to tumor necrosis factor alpha (TNF-a), suitably a disease selected from rheumatoid arthritis, rheumatoid arthritis juvenile, psoriatic arthritis, plaque psoriasis and / or ankylosing spondylitis;

as defined elsewhere herein.

Liquid pharmaceutical compositions are suitably administered parenterally, suitably by subcutaneous injection.

Examples

Materials and equipment

The following samples were used in (or to produce) the Examples and Screening Experiments described herein:

All excipients and ingredients used in the preparation of the Example formulations described herein were readily available commercially.

The following kits were used in the Screening Examples and Experiments described herein:

Analytical techniques and protocols

The individual protocols for each of the above analytical methods are successively described below, and these protocols were used in the references in the Examples and Screening Experiments to those analytical methods.

Analytical sample preparation

The analytical methods indicated above, and for which the protocols are provided below, were carried out on samples of each of the exemplified formulations, and which are described in more detail below, which were exposed to various stability tests. Samples of each of these formulations were subjected to: T ⁰ (samples not subjected to any stress), T ^mec (samples subjected to mechanical stress), T ^heat (samples subjected to heat stress), T ^{1 month} (samples stored at temperature environment for 1 month).

After preparation, the formulations were loaded into the syringes. In the T ^mec mechanical stress test, the syringes were shaken horizontally for 2 hours on an IKA KS 4000ic control shaker at 25 ± 3 ° C and 400 rpm. In the heat T ^heat stress test, the syringes were placed for 1 week at 40 ° C. In the T ^{1 month} storage test, the syringes were stored for 1 month at 25 ° C. In the storage test T ^3months, the syringes were stored for 3 months at 25 ° C.

A1 - High Performance Size Exclusion Chromatography ( HP-SEC)

The HP-SEC was performed according to the following protocols and parameters:

A2 - Hydrophobic Interaction Chromatography ( HIC)

HIC analysis of formulations 1-4 and the control formulation was performed on an HP1100 system equipped with a UV detector (Agilent Technology, USA).

Before injection, the -20 mg / ml etanercept material was diluted 1 1 with sample dilution buffer containing 25 mM sodium phosphate pH 6.3 at -10 mg / ml. Subsequently, a 1 1 mixture was made with Eluent A (binding buffer composed of 0.1 M sodium citrate and 0.8 M sodium sulfate, pH 6.0) to a final concentration of ~ 5 mg / ml.

The following analysis parameters were used:

A3 - Visual inspection

For visual inspection, the syringes were inspected for the presence or absence of visible particles under gentle manual radial shaking for 5 seconds against a white background and for 5 seconds against a black background according to the European Pharmacopoeia. The inspection was carried out by two independent examiners. To further classify the particle content, the method described in the "Deutscher Arzneimittel-Codex" (DAC) was used. The classification can be described as follows:

A4 - UV-Vis Spectroscopy

UV spectroscopy was performed using a Tecan Safire2 plate reader on 96-well plates at 280 nm. For each sample, 3 wells (n = 3) were measured, each with 200 µl of solution at a concentration of 1 mg / ml, to minimize measurement errors. After measurement, all data was subtracted by blank spectrum and corrected for path length (A998nm / A900nm). The protein concentration was calculated using a specific absorbance coefficient of 1.13 at 280 nm for a protein concentration of 1.0 mg / ml at a path length of 1 cm. The concentration of etanercept was calculated using the following formula:

A280nm = £ 280nm X C X d

Where:

At ²⁸⁰ nm = absorbance measured at 280 nm

£ ²⁸⁰ nm = extinction coefficient at 280 nm in ml mg-1 cm-1

(£ ²⁸⁰ nm = 1.13 ml mg-1 cm-1)

C = concentration in mg / ml

d = path length in cm

A5 - SDS-PAGE

SDS-PAGE was performed as follows.

Briefly, for non-reducing conditions 40 µl of an etanercept sample (~ 1.0 mg / ml) was mixed with 50 µl of Tris Glycine sample buffer (2x) and 10 µl of 10 mM maleimide solution. For reducing conditions, 40 µl of an etanercept sample (~ 1.0 mg / ml) were mixed with 50 µl of T ris Glycine sample buffer (2x) and 10 µl of a 100 mg / ml DTT solution. Both samples were incubated on a heating block (Thermo Mixer Compact, Eppendorf AG, Germany) at 85 ° C for 4 minutes prior to gel loading. After heating, the samples were cooled to RT and then 12.5 µl of sample was loaded into the respective wells of Novex 4-20% Tris-Glycine gels (1mm x 12 wells, Life Technologies, USA). The gels were run at a constant current of 30 mA for -60 minutes in an XCell SureLock Mini-Cell (Invitrogen, USA) in combination with a PowerPac 200 power supply (BioRad, USA). A 10 µl aliquot of previously stained protein standard SeeBlue Plus2 (Life Technologies, USA) was used as a molecular weight marker.

After the gels were processed, they were immediately released from the cassettes and placed in 50 ml of Simply Blue Stain Solution (Life Technologies, USA). Staining was performed on a staining tray on a horizontal shaker (GFL 3012, Gesellschaft für Labortechnik, Germany) overnight. The gels were discolored with highly purified water for a minimum of 3 hours, including several washing steps (3 rinses for 5 minutes with 50 ml of highly purified water and 5 rinses for 15 minutes with 50 ml of highly purified water) .

Example 1: general methods of preparation of formulations

The commercial reference product Enbrel® (RMP, formulated at 50 mg / ml, 0.5 ml per syringe) was used to prepare all the formulations described in this Example.

Material was drawn from the syringes into a sterile PETG bottle under laminar flow conditions by gently pushing the plungers. After grouping, two examiners performed a visual inspection.

Four aliquots were prepared from the pooled bulk solution by transferring the material to sterile PETG bottles. The bottles have been placed at 2-8 ° C for later phases of the project. 14 ml of the pooled solution were used for the formulations. In addition, 500 µl of the pooled material was transferred to a polypropylene tube (Eppendorf) and stored at -80 ° C for possible analytical purposes.

The RMP material was dialyzed in an amino acid free buffer containing 25 mM phosphate, 100 mM sodium chloride and 1% w / v sucrose with pH 6.3. Dialysis was performed at 4 ° C using two slide-a-lyzer dialysis cassettes (Thermo Scientific) with a molecular weight limit of 10 kDa and up to 12 ml capacity. Subsequently, both dialysed etanercept fractions were combined in a 15 ml PP tube without particles (Greiner bio-one). Subsequently, two examiners carried out a visual inspection.

In addition, two additional dialysis preparations (i) and (ii) were performed overnight at 4 ° C in slide-a-lyzer dialysis cassettes (Thermo Scientific) with a molecular weight limit of 10 kDa and up to 12 ml of ability.

For these additional dialyses the following buffers were prepared:

i) Citric acid 25 mM

50 mM sodium chloride

pH 6.3

ii) 25 mM phosphate

50 mM sodium chloride

pH 6.3

After dialysis, the protein concentration was determined by 280 nm UV spectroscopy at a 1: 50 dilution.

i) 43.4 mg / ml; ii) 38.6 mg / ml

The determined protein concentrations were taken into account during the preparation of the final formulations. The formulations were transferred to 15 ml PP tubes without particles (Greiner bio-one).

In 1 ml siliconized syringes (BD Hypak SCF combined with a 29G needle) 520 µl of placebo or of the pooled RMP were manually introduced at ~ 20 mg / ml under laminar air flow conditions. Particle-free pipette tips (SafeSeal-Tips Professional, Biozym, Germany) were used to transfer the materials to the syringes. The entry of air bubbles was prevented by adding the liquid slowly to the syringe on the wall. The syringes were sealed with plungers (HYPAK b Sc F 1MLL W4023 FLUR DAIKYO LID) using a Bausch & Strobel SVP100 insertion and closure machine. The emboli were systematically placed in the same position so that considerable headspace was created within the syringes, allowing air-liquid interactions during mechanical stress. The above-mentioned procedures were employed in the preparation of the example formulations detailed in Table 1.

Table 1 - Example formulations

Formulation 1

The dialyzed Enbrel RMP material was diluted to -20 mg / ml using sterile filtered buffer consisting of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose pH 6.3. No pH adjustment was required. This produced a final aqueous liquid formulation containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v

with pH 6.3.

Formulation 2

The RMP Enbrel material was dialyzed using sterile filtered buffer composed of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose with pH 6.3 to finally produce -40 mg / ml of etanercept in said buffer solution. A separate 50 mM aspartate / aspartic acid solution, pH 6.3, was formed within the same sterile filtered buffer described above. This 50 mM aspartate / aspartic acid solution was then added (RMP solution: amino acids in 1: 1 dilution) to the RMP solution to obtain a final aspartate / aspartic acid concentration of 25 mM and a final RMP concentration of ~ 20 mg / ml. No pH adjustment was required. This produced a final aqueous liquid formulation containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM aspartic acid;

with pH 6.3.

Formulation 3

The dialyzed Enbrel RMP material from dialysis (i) was diluted to -20 mg / ml using sterile filtered buffer to a final composition of 25 mM citrate, 100 mM NaCl, 1% sucrose and 25 mM histidine with pH 6, 3. This produced a final aqueous liquid formulation containing:

- 20 mg / ml of etanercept;

- 25 mM citrate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM histidine;

with pH 6.3.

Formulation 4

The dialyzed Enbrel RMP material from dialysis (ii) was diluted to -20 mg / ml using sterile filtered buffer to a final composition of 25 mM phosphate, 50 mM NaCl, 4% sucrose and 12.5 mM lysine proline 12 , 5 mM with pH 6.3. This produced a final aqueous liquid formulation containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 50 mM NaCl;

- sucrose at 4% w / v;

- 12.5 mM lysine;

- 12.5 mM proline;

with pH 6.3.

Control formulation

The dialyzed Enbrel RMP material was diluted to -40 mg / ml using sterile filtered buffer consisting of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose pH 6.3. A 50 mM L-Arg HCl solution, pH 6.3 (RMP solution: amino acids in 1: 1 dilution) was added to the RMP solution to obtain a final amino acid concentration of 25 mM and a final RMP concentration of -20 mg / ml. No pH adjustment was required. This produced a final aqueous liquid formulation containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM L-arginine hydrochloride;

with pH 6.3.

Example 2 - Formulation analysis and stability studies

The formulations in Table 1 were subjected to analysis by analytical methods A1-A5 described above. The results of these analytical tests are presented below in tabulated form. Each test result is refers to 1 of 4 samples selected from T ^o (samples not subjected to any stress), T ^mec (samples subjected to mechanical stress), T ^heat (samples subjected to heat stress), T ^{1 month} (samples stored at room temperature for 1 month).

A1 - High Performance Size Exclusion Chromatography Results ( HP-SEC)

Table 2 - High Performance Size Exclusion Chromatography (HP-SEC)

A2 - Results of hydrophobic interaction chromatography ( HIC)

Table 3 - Analysis by hydrophobic interaction chromatography (HIC)

A3 - Visual inspection results

T a b la 4 - R e s u l ta d o s de la in s p e c tio n v is u a l

A4 - Results of UV-Vis spectroscopy

T a b la 5 - R e s u l ta d o s de l a n á l is e s p e c t r o s c o p ic o U V - V is

A5 - SDS-PAGE Results

Table 6 - Results of the SDS-PAGE analysis

Example 3: general methods of preparation of formulations

The commercial reference product Enbrel® (RMP, formulated at 50 mg / ml, 0.5 ml per syringe) was used to prepare the control formulation described in this Example.

This RMP material for the control formulation was pooled in a 15 ml Falcon tube under laminar air flow conditions. Pooling was done gently through the primary needle attached to the RMP syringes, to avoid the formation of air bubbles. The pooled RMP material was dialyzed against a buffer containing 25 mM arginine-HCl, 25 mM phosphate, 100 mM sodium chloride, and 1% w / v sucrose pH 6.3. Dialysis was performed at 2-8 ° C, using slide-a-lyzer dialysis cassettes (Thermo Scientific) with a molecular weight limit of 10 kDa and up to 12 ml capacity. The dialyzed RMP material was filtered (sterile) using 0.22 pm PVDF syringe filters. The final concentration of Enbrel® in the control formulation was adjusted to 20 mg / ml and 50 mg / ml using a buffer containing 25 mM arginine-HCl, 25 mM phosphate, 100 mM sodium chloride and 1% w / w sucrose. v with pH 6.3.

In addition, an etanercept drug substance other than the RMP material was used to prepare all other formulations in this Example. First, a solution of the drug etanercept was concentrated using an Amicon Ultra-15 centrifugal concentrator with a molecular weight limit of 30 kDa to obtain an aqueous solution containing etanercept at a concentration of approximately 70 mg / ml.

The above-mentioned procedures were employed for the preparation of the example formulations detailed in Table 7.

Table 7 - Example formulations (each in a protein concentration of 20 and 50 mg / ml)

Formulation A

The aqueous solution of etanercept material was dialyzed using a sterile filtered buffer composed of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose, and 25 mM L-aspartic acid with pH 6.3 to finally yield 55.7 mg / ml of etanercept in said buffer solution. The dialyzed etanercept solution thus obtained was diluted with the same sterile filtered buffer to result in a liquid formulation containing 20 mg / ml of etanercept and a liquid formulation containing 50 mg / ml of etanercept. This produced a final aqueous liquid formulation (Formulation A - 20 mg / ml) containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM aspartic acid;

with pH 6.3,

and a final aqueous liquid formulation (Formulation A - 50 mg / ml) containing:

- 50 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM aspartic acid;

with pH 6.3.

Formulation B

The aqueous solution of etanercept material was dialyzed using sterile filtered buffer composed of 25 mM phosphate, 50 mM NaCl, 4% w / v sucrose, 12.5 mM L-lysine HCl and 12.5 mM L-proline with pH 6 , 3 to finally produce 63.9 mg / ml of etanercept in said buffer solution. The dialyzed solution of etanercept thus obtained was diluted with the same sterile filtered buffer to result in a liquid formulation containing 20 mg / ml of etanercept and a liquid formulation containing 50 mg / ml of etanercept. This produced a final aqueous liquid formulation (Formulation B - 20 mg / ml) containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 50 mM NaCl;

- sucrose at 4% w / v;

- 12.5 mM lysine;

- 12.5 mM proline;

with pH 6.3,

and a final aqueous liquid formulation (Formulation B - 50 mg / ml) containing:

- 50 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 12.5 mM lysine;

- 12.5 mM proline;

with pH 6.3

Formulation C

The aqueous solution of etanercept material was dialyzed using a sterile filtered buffer composed of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose with pH 6.3 to finally produce 60.5 mg / ml of etanercept in said solution. tampon. The dialyzed etanercept solution thus obtained was diluted with the same sterile filtered buffer to result in a liquid formulation containing 20 mg / ml of etanercept and a liquid formulation containing 50 mg / ml of etanercept. This produced a final aqueous liquid formulation (Formulation C - 20 mg / ml) containing: - 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

. 100 mM NaCl;

- sucrose at 1% w / v;

with pH 6.3,

and a final aqueous liquid formulation (Formulation C - 50 mg / ml) containing:

- 50 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

with pH 6.3.

Formulation D

The aqueous solution of the etanercept material was dialyzed using a sterile filtered buffer composed of 25 mM citrate, 100 mM NaCl, 1% w / v sucrose, and 25 mM L-histidine with pH 6.3 to finally yield 59.8 mg / ml of etanercept in said buffer solution. The dialyzed etanercept solution thus obtained was diluted with the same sterile filtered buffer to result in a liquid formulation containing 20 mg / ml of etanercept and a liquid formulation containing 50 mg / ml of etanercept. This produced a final aqueous liquid formulation (Formulation D - 20 mg / ml) containing:

- 20 mg / ml of etanercept;

- 25 mM citrate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM histidine;

with pH 6.3,

and a final aqueous liquid formulation (Formulation D - 50 mg / ml) containing:

- 50 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM histidine;

with pH 6.3.

Control formulation

A dialyzed Enbrel RMP material was further diluted to -60 mg / ml in sterile filtered buffer consisting of 25 mM phosphate, 100 mM NaCl, 1% w / v sucrose and 25 mM L-arginine HCl pH 6.3. This produced a final aqueous liquid formulation (Control - 20 mg / ml) containing:

- 20 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM L-arginine hydrochloride;

with pH 6.3,

and a final aqueous liquid formulation (Control - 50 mg / ml) containing:

- 50 mg / ml of etanercept;

- 25 mM phosphate buffer;

- 100 mM NaCl;

- sucrose at 1% w / v;

- 25 mM L-arginine hydrochloride;

with pH 6.3.

510 µl of control (RMP material) or of the drug substance etanercept at 20 mg / ml or 50 mg / ml under air flow conditions were manually inserted into 1 ml siliconized syringes (BD Hypak SCF combined with a 29G needle). laminate. Particle-free pipette tips (SafeSeal-Tips Professional, Biozym, Germany) were used to transfer the materials to the syringes. The entry of air bubbles was prevented by adding the liquid slowly to the syringe on the wall. Syringes were sealed with plungers (HYPAK BSCF 1MLL W4023 FLUR DAIKYO LID) using a Bausch & Strobel SVP100 insertion and closure machine. The emboli were systematically placed in the same position so that considerable headspace was created within the syringes, allowing air-liquid interactions during mechanical stress.

Example 4 - Formulation analysis and stability studies

The formulations in Table 7 were subjected to analysis by analytical methods A1-A5 described above. The results of these analytical tests are presented below in tabulated form. Each test result refers to 1 of 5 samples selected from T ⁰ (samples not subjected to any stress), T ^mec (samples subjected to mechanical stress), T ^heat (samples subjected to heat stress), T ^{1 month} (samples stored at room temperature for 1 month), and T ^3months (samples stored at room temperature for 3 months).

A1 - High Performance Size Exclusion Chromatography Results ( HP-SEC)

Table 8 - High performance size exclusion chromatography (HP-SEC) with formulations containing 20 mg / ml of etanercept.

Table 9 - High performance size exclusion chromatography (HP-SEC) with formulations containing 50 mg / ml of etanercept.

Table 10 - HP-SEC / SDS-PAGE monomer content *

A2 - Results of hydrophobic interaction chromatography ( HIC)

Table 11 - Analysis by hydrophobic interaction chromatography (HIC) with formulations containing 20 mg / ml of etanercept

Table 12 - Analysis by hydrophobic interaction chromatography (HIC) with formulations containing 50 mg / ml of etanercept

A3 - Visual inspection results

Table 13 - Visual inspection results

A5 - SDS-PAGE Results

Table 14 - Results of the SDS-PAGE analysis

List of abbreviations:

To absorbance

Arg arginine

Asp aspartic acid

Glu glutamic acid

His histidine

Lys lysine

Pro proline

AUC area under the curve

cIEF capillary isoelectric focusing

DLS dynamic light scattering

DS drug substance

pDSC Differential Scanning Calorimetry

DSF dynamic scanning fluorimetry

FNU nephelometric unit of formazin

HIC hydrophobic interaction chromatography

HP-SEC High Pressure Size Exclusion Chromatography MFI Microflow Imaging

OD optical density

PDI polydispersity index

PETG polyethylene terephthalate glycol

PFS pre-filled syringes

Ph.Eur. European Pharmacopoeia

PP polypropylene

RMM resonant mass measurement

RMP commercial reference product (Enbrel®)

SDS-PAGE sodium dodecyl polyacrylamide gel electrophoresis Tm melting temperature

UV ultraviolet

Claims (12)

1. A liquid pharmaceutical composition comprising: 20 to about 70 mg / ml of etanercept (including any biosimilars thereof); a 10 to 40 mM buffer system, the buffer system being a phosphate buffer system and / or a citrate buffer system; 20 to 150 mM sodium chloride; 0.5 to 5% by weight of sucrose; Y either aspartic acid, or a combination of lysine and proline; where: the liquid pharmaceutical composition has a pH between 6.1 and 6.5; the liquid pharmaceutical composition is free of arginine or comprises arginine in a concentration of at most 0.1 mM; the liquid pharmaceutical composition is free of sulfur-containing amino acid (s) or comprises sulfur-containing amino acid (s) in a concentration of at most 0.1 mM; the liquid pharmaceutical product is free of nitrogen-containing chelating agent (s) or comprises nitrogen-containing chelating agent (s) in a concentration of at most 0.1 mM; Y the liquid pharmaceutical composition containing a combination of lysine and proline contains 3.5 to 4.5% by weight of sucrose. 2. The liquid pharmaceutical composition according to claim 1, wherein, when the liquid pharmaceutical composition comprises both lysine and proline, the molar ratio of lysine to proline is between 2: 1 and 1: 2. 3. The liquid pharmaceutical composition according to claim 1 or 2, comprising lysine in combination with proline, in a concentration (collective) of 10 to 40 mM. 4. The liquid pharmaceutical composition according to any of the preceding claims, comprising a phosphate buffer system. 5. The liquid pharmaceutical composition according to any of the preceding claims, wherein the molar concentrations of sucrose and sodium chloride are interrelated by Equation (1): [stabilizer] ^moi * [tonicity regulator] ^moi = A ^moi * 2n Equation (1) where [stabilizer] ^{m oi} is the molar concentration of sucrose; [tonicity regulator] ^{m oi} is the molar concentration of sodium chloride; A ^moi is a reference molar concentration constant; and n is any positive or negative number; where A ^moi is between 2,000 and 4,000 mM2; and n is between -2 and 2. 6. The liquid pharmaceutical composition according to any of the preceding claims, wherein the liquid pharmaceutical composition comprises: - 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept; - 20-30 mM buffer system selected from a phosphate buffer system or a citrate buffer system; - 40-120 mM sodium chloride (suitably 40-60 mM or 90-110 mM); - 0.5-5.0% by weight of sucrose (suitably 0.5 to 1.5% by weight or 3.5 to 4.5% by weight); Y - water (for injection); wherein the composition has a pH between pH 6.1 and 6.5; Y wherein the composition comprises: - 20-30 mM aspartic acid; or - a combination of 10-15 mM lysine and 10-15 mM proline; Y wherein the composition is free of arginine or comprises arginine in a concentration of at most 0.1 mM; wherein the composition is free of sulfur-containing amino acid (s) or comprises sulfur-containing amino acid (s) in a concentration of at most 0.1 mM; wherein the composition is free of nonionic surfactants or comprises one or more of said surfactants in a (collective) concentration of at most 0.1 mM; Y wherein the composition is free of nitrogen-containing chelating agent (s) or comprises nitrogen-containing chelating agent (s) in a concentration of at most 0.1 mM. 7. The liquid pharmaceutical composition according to any of the preceding claims, wherein the liquid pharmaceutical composition consists essentially of: - 40-60 mg / ml (or alternatively 20-30 mg / ml) of etanercept; - 20-30 mM buffer system selected from a phosphate buffer system or a citrate buffer system; - 40-120 mM sodium chloride; - 0.5-5.0% by weight of sucrose; Y - water (for injection); wherein the composition has a pH between pH 6.2 and 6.4; and wherein the composition comprises: - 20-30 mM aspartic acid; or - a combination of 10-15 mM lysine and 10-15 mM proline. 8. The liquid pharmaceutical composition according to any of the preceding claims, wherein the liquid pharmaceutical composition consists essentially of: - 50 mg / ml (or alternatively 25 mg / ml) of etanercept; - 25 mM phosphate buffer system; - 100 mM sodium chloride; - sucrose at 1% by weight (or 30 mM / -1 mM); - 25 mM aspartic acid; Y - water (for injection); wherein the composition has a pH of 6.3. 9. The liquid pharmaceutical composition according to any of the preceding claims, wherein the liquid pharmaceutical composition consists essentially of: - 50 mg / ml (or alternatively 25 mg / ml) of etanercept; - 25 mM phosphate buffer system; - 50 mM sodium chloride; - 4% by weight (or 120 mM / -4 mM) of sucrose; - 12.5 mM lysine; - 12.5 mM proline; Y - water (for injection); wherein the composition has a pH of 6.3. 10. A drug delivery device (eg, syringe or pre-filled pen, or intravenous bag) comprising a liquid pharmaceutical composition according to any of claims 1 to 9. 11. A liquid pharmaceutical composition according to any of claims 1 to 9 for use in therapy. 12. A liquid pharmaceutical composition claimed in any one of claims 1 to 9 for use in the treatment of rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, plaque psoriasis and / or ankylosing spondylitis.